• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.43-49
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• 2001

This study was performed to elucidate the effects of copper poisoning on the reproductive organ of rats. After consecutive oral administrations of copper sulfate, the weights of testis, the numbers and motilities of sperms, organ weights and histological changes of testes were compared between control and experimental groups. 1. Testis weights of 1,000, 2,000 or 4,000 ppm/kg of copper sulfate-administrated rats gradually decreased compared with control group, and the values no significant changes. 2. The sperm numbers of 1,000, 2,000 or 4,000 ppm/kg of copper sulfate-administrated rats were lowered in dose dependent manners than those of control group's and the values no significant changes. 3, The motilities sperms of 1,000, 2,000 or 4:000 ppm/kg of copper sulfate-administrated rats decreased significantly in dose-dependent manners compared with those of control group's. 4. The weights of livers and kidneys of 1,000, 2,000 or 4,000 ppm/kg of copper sulfate-administration rats decreased or increased. 5. Necrosis of hepatocytes around the central veins and infiltrations of fine granules-harboring macrophages in periportal and interstitial tissues were found out in the livers of copper sulfate-administrated rats. The Bowman's capsule and tubules of kidneys were filled with hyaline material.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.2
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• pp.193-199
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• 2015

We observed that the growth and physiological change in Haliotis discus discus by low-dose irradiation with gamma ray.Irradiation with gamma ray was undertaken by using the low-level irradiation facility ($^{60}CO$) in cooperation with the Institute of Nuclear Science and Technology at Jeju National University. The parent abalones were attached one by one and then fixed by using the rubber band to the front side of the fabricated case for irradiation with gamma ray. The experimental plots of irradiation with gamma ray were set as 10, 15, 20 and 25 Gy and the 25 female abalones and 10 male abalones were utilized for each experimental plot. The sperms and eggs were fertilized by setting an interval for each dose to prevent mixing with other experimental plots when fertilizing the sperms and eggs for each dose of irradiation with gamma ray. As for the fertility, it was confirmed to be 85% the control and 10 Gy groups, whereas it was found to be 80%, 65% and 50% in the 15 Gy, 20 Gy and 25 Gy groups, respectively. As a conclusion, the hatching rate and attachment rate were higher at 10 and 15 Gy than the other experimental plots, and the growth rate was higher at 20 Gy than the other experimental plots. Also the changes in lysozyme activity in accordance with the stress of water temperature were found to have a significant increase in the other experimental plots as compared with the control plot at the end of 0 h. The changes in lysozyme activity have remained constant in all the experimental plots at the end of 12 h. These results allowed us to confirm that lysozyme was undertaking the biodefense action by reacting sensitively to the stress of water temperature in the control experimental plot. As for the other experimental plots, they are believed to avoid the biodefense mechanism due to the high degree of anti-parasite mechanism and anti-viral mechanism. Thus, it is believed that it would be imperative to conduct development and research on breeds that were potent for environmental tolerance by applying the method of irradiation with gamma ray to other marine animals and plants.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1527-1533
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• 2001

Garole is a prolific, rare, less known and small size Indian sheep breed found in low and humid Sunderban region of West Bengal. Although information on stored Garole ram liquid semen upto 24 h is available, but there is a need to further investigate the short-term and long-term preservability of Garole ram semen for extensive utilization of this valuable germplasm by artificial insemination. The aim of the present study was to apply computer-assisted sperm analysis technique for assessing the motion characteristics of Garole ram semen stored (i) in liquid state at refrigeration temperature for short-term preservation upto 48 h and (ii) in frozen state at $-196^{\circ}C$ for long-term preservation after packaging in mini straws. Short-term preservation had a significant effect on motility (p<0.01) as the motility progressively decreased from 90.1% at 0 h to 85.5% and 73.2% after 24 and 48 h of storage, respectively. Although the decline in rapid moving sperms was also significant (p<0.01) on storage but the decrease was more pronounced at 48 h as compared to 24 h of storage period. Storage of chilled semen had also a significant effect on % linearity (p<0.05), % straightness (p<0.01), sperm velocities (p<0.01), amplitude of lateral head displacement (p<0.01) and beat frequency (pO.Ol) of spermatozoa. The replication had a significant effect for all the variables except average path and straight line velocity. However, the interactions of short-term storage and replication were non-significant for most of the variables except % of medium moving sperms, sperm velocities and beat frequency. On long-term preservation of Garole ram spermatozoa under controlled conditions the mean post-thaw recovery of 70.4 and 71.4% motile spermatozoa was achieved having 48.8 and 48.9% of rapidly motile spermatozoa, respectively in both the replicates. The effect of replication on cryopreservation was significant (p<0.05) on amplitude of lateral head displacement and beat frequency, but there was no significant effect on motility, rapidly motile spermatozoa, linearity, straightness and sperm velocities of frozen-thawed spermatozoa. It can be concluded from these results that an average 70% motility can be achieved on storage of Garole ram semen in chilled liquid state upto 48 h or in liquid nitrogen after freezing under controlled conditions in straws. However, further studies are required to evaluate the fertility of short-term and long-term preserved Garole ram semen for extensive use of this prolific sheep breed.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.11 no.2
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• pp.59-67
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• 1984

In vitro fertilization have been performed to know whether the frozen semen has fertilizing ability and can be used clinically. The results of cultured and developed embryos obtained are as follows: 1. The semen was frozen in three media for the good viability. The viability was more than 50% and the motility was also moderate (grade III), 2. As the 33 oocytes were collected from 45 follicles, the oocyte recovery rate was 73.3%. Among them, mature and immature ova were 5% each, and premature ova were 69.7%, When the first polar body was appeared, above ova were inseminated after adequate incubation with activated sperms. 3. The main components of three freezing medium containing egg yolk, glycerol and pyruvate respectively were the best for sperm viability, and Ham's F-10 medium was used for the fertilization and culture of eggs. 4. The results of in vitro fertilization of 33 ova, showed the second polar body developed in 12%, polyspermia in 24%, 1-cell embryo in 21% and 2-cell embryo in 9%. One mature ova developed to blastocyst via 16-cell to 32-cell embryo. The fertilization rate was 66%. 5. Above mentioned results represent that the frozen semen has fertilizing ability and can be used practically in the clinic.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.3
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• pp.439-447
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• 1992

Eight healthy Holstein bulls, 4-6 years old were used to study the effect of season of the year on the biophysical characteristics of semen. Semen was collected twice a week by AV (artificial vagina) over one-year period. The analyses revealed that all the basic seminal traits studied were differed significantly due to season, except the ejaculate volume and consistency and the percentage of swollen spermatozoa in a hypo-osmotic fructose-citrate solution. Ejaculates collected during hot summer season had significantly lower sperm motility, concentration and total counts, and higher percentage of dead spermatozoa than those collected during winter time. Warm spring had moderate semen quality. The temperature-humidity index was calculated and it was associated (p < 0.01) negatively with the ejaculate pH, sperm concentration and total counts, and positively with the % of dead sperms. Ejaculate volume, percentage of swollen spermatozoa, individual motilities did not correlate significantly with the change in temperature-humidity index values. The total live, motile spermatozoa per ejaculate during both the winter and spring seasons showed significant increase of about 37% and 32% respectively over the summer season. Also, rectal temperatures of the bulls were elevated during the hot summer season, while the values of blood hemoglobin and packed-cell volume were decreased.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.15-21
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• 1999

The aim of the present study was to determine the effect of paternal sex chromosome on early development of buffalo embryos fertilized and cultured in vitro. Embryos were produced in vitro from abattoir derived buffalo oocytes. The cleaved embryos were cocultured with buffalo oviductal epithelial cells and evaluated on day 7 under the phase contrast microscope to classify development. The embryos which reached the morula/blastocyst stage were fast developing, the embryos which were at 16-32 cell stage were medium developing and the embryos below 16 cell stage were slow developing. The embryos which showed some fragmentation in the blastomeres or degenerated blastomeres, were degenerating. Sex of emberyos (n=159) was determined using PCR for amplification of a male specific BRY. 1 (301 bp) and a buffalo specific satellite DNA (216 bp) fragments. The results thus obtained show that 1) X and Y chromosome bearing sperms fertilize oocytes to give almost equal numbers of cleaved XX and XY embryos, 2) male embryos develop faster than female embryos to reach advanced stage and 3) degeneration of buffalo embryos is not linked with the paternal sex chromosome. We suggest that faster development of males is due to differential processing of X and Y chromosome within the zygote for its activation and / or differential expression of genes on paternal sex chromosome sex chromosome during development of buffalo embryos fertilized and cultured in vitro which may be attributed to a combination of genetic and environmental factors.

• Environmental Analysis Health and Toxicology
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• v.18 no.2
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• pp.77-87
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• 2003

성게의 정자를 이용한 생물검정은 해양환경의 독성평가에 널리 사용되고 있다. 그러나, 국내산 성게류의 경우 산란기가 3~11월로 겨울에는 생물검정 실칩이 불가능한 단점이 있다. 이를 보완하기 위하여 겨울에 산란이 가능한 아무르불가사리(Arterias amurensis)를 생물검정용으로 개발하였다. 불가사리류의 생식과 발생과정은 성게류와 유사하므로 성게류를 이용한 실험 방법을 아무르불가사리에 적용할 수 있다. 그러나, 생물검정 실험 결과의 유효성은 실험조건에 의하여 달라질 수 있으므로, 본 연구에서는 실험 결과에 큰 영향을 미칠 수 있는 조건들 즉. 수정막의 발달 시간, 정자와 알의 비율, 정자의 활력이 유지되는 시간, 정자의 노출 시간, 그리고 시료의 염분 범위 등을 최적화 하고자 하였다. 수정막의 발달이 완전하게 이루어지는 최소 시간은 60분이었고, 시료에 주입할 정자와 알의 비율은 3,000이상일 때 수정률이 높게 유지되었다(70%이상). 정자의 활력은 해수에 희석한 후 3시간 이상이 경과하면 급격히 감소하였다. 정자의 노출시간은 20~60분의 범위에서 구리에 대한 $EC_{50}$이 크게 달라지지 않았다(변동계수: 17%).실험에 적절한 시료의 염분 범위는 26~38 psu이었다. 아무르불가사리 정자의 민감도는, 일반적으로 해양환경 평가에 흔히 사용되는 생물들의 민감도 범위의 중간 정도에 해당하였다. 따라서 아무르불가사리의 정자는 해양환경 평가를 위한 생물검정용으로 적합한 것으로 여겨진다.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.1
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• pp.1-7
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• 2009

Quality of seed in the mulberry silkworm, Bombyx mori (L.) is determined by many important factors, wherein unfertilized eggs play an important role. Unfertilization of eggs are caused by several reasons such as, abnormality in the sexual organs of the male and female, abnormal development of the micropylar end of the egg, unfavorable environmental conditions during spinning, cocoon preservation, imperfect handling of moths, mating, ovipostion, cold storing of pupae / moths and indiscriminate use of male moths etc. Though the presence of unfertilized eggs would in no way affect the fertilized ones and their quality directly, the frequency of their occurrence underrates the quality and brings down the hatching percentage. Lower the occurrence of unfertilized eggs, higher is the rating of seed quality. Of the various intrinsic and extrinsic factors and events involved in egg deposition of an adult silk moth, mating is an instinct and a biological obligation for the ultimate perpetuation of the species and a must to provide stimulus for oogenesis and bring about biochemical changes in the spermatophore of the silkworm in order to ensure the presence of sufficient number of normal sperms and testicular fluid in the female reproductive organ, activating ovulation and accelerating oviposition behavior and egg deposition. An attempt has been made in this article to briefly elucidate the characteristics of unfertilized eggs, causes of their occurrence and its impact as well as the significance in silkworm seed production.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.287-296
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• 2011

Mammalian fertilization is a complex cascade process consisting of sperm migration through the female reproductive tract, physiological changes to sperm such as sperm capacitation and acrosome reaction, and sperm-egg interaction in the oviduct in vivo. On the other hand, in vitro fertilization (IVF) is a process by which egg cells are fertilized by sperm outside the body: in vitro. IVF has been used for a variety of purposes in reproductive biotechnology for human and animals. The discovery of sperm capacitation in 1951 promoted the development of IVF technology. In the initial stage of IVF, sperm capacitation in preincubation medium was shown to be essential to fuse with eggs. Besides, sperms should detour some of the in vivo regulations for IVF. This review introduces a general mammalian fertilization process, including sperm capacitation, removal of cumulus matrix, acrosome reaction, and sperm-egg fusion and focuses on the roles of key biochemical molecules, signal mechanisms, and genes involved during IVF and novel results of sperm-oocyte interaction elucidated in various gene-knockout mice models.