• Journal of Fisheries and Marine Sciences Education
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• v.26 no.6
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• pp.1402-1411
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• 2014

This study about morphological observation of pre- or post ovulated eggs obtained from artificially maturated female eels. Female eels were divided with two groups as wild eels from nature and feminized eels from farm. Artificial maturation had been conducted with the established methods in this laboratory, and then maturate eggs sampled at fixed 3 times and these were observed with stereomicroscope. Rate of increased body weight (RIW) were measured with 2 times. Egg diameters and development of oil droplets are determined for standardization of egg maturation degree, and the transparency of egg cytoplasm and the homogeneous degrees of egg size were referred to determine of egg quality. Rate of increased body weight (RIW) were good in range about 10 % at final salmon pituitary extracts (SPE) injection time and in range about 20 % at $17{\alpha}$, $20{\beta}$-dihydroxy-4-pregnen-3-one (DHP) injection time. Good matured egg for fertilization had $900-1000{\mu}m$ diameter, and they had about 50 oil droplets in size about $100{\mu}m$ diameter. There were not predominant differences at egg size and oil droplet development between wild female and feminized eels.

• Journal of Aquaculture
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• v.4 no.1
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• pp.31-66
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• 1991

This paper deals with the reproductive ecology e.g., number of the pre-spawning moults, morphological characteristics of the pre-spawning moult the common moult, daily ration druing a molting cycle mating behavior, structures of spermatozoa and spermatophore, structure of vas deferens, mechanisms of the oviposition and brooding into the egg-chambers, a suitable time for the artificial mating and fertilization, time sequence of the oviposition and brooding into egg-chambers from the copulation, responses to temperature and chlorinity on the egg development and hatching, effect of temperatures on duration of egg development, physical mechanism of the egg hatching, to make an attempt for the artificial spawning and brooding to establish a suitable system of the artificial seedling-production for the aquaculture. 1. Females molted commonly $8{\~}10$ times at an interval of $17{\~}18$ days at $28^{\circ}C,\;3.26\~4.35\%_{\circ}$ while the prespawning moltings were $4{\~}5$ times at an interval of $13{\~}14$ days. The suitable state for artificial copulation was within 14 hours elapsed from the prespawning moltings (most suitable state was within 8 hours). Males discharged a gelatinous spermatophore and placed it on the females sternum during copulation. Oviposition was seen $6{\~}17$ hours after copulation. External fertilization was considered to take place at oviposition. Fertilized eggs held in egg-chambers forming between pleopods were about $5000{\~}6000$ in females those sizes about 6.5 cm in body length. 2. Eggs immediately after oviposition were elliptic shape, measuring $0.58{\times}0.48$ mm up to hatching. Their sizes increased with egg development and finally reached $0.85{\times}0.54$ mm up to hatching. The relationship between the long axis of the egg(Y in U) and days elapsed(X) was expressed as Y= 5.60194 + 0.007358X. The eggs performed superficial cleavage and their cleavage furrows became visible at the 4-daughter-nucleus stage. The eggs showed normal development up to hatching at water temperature range of $22{\~}30^{\circ}C$ (optimum temperature : $26{\~}28^{\circ}C$) and at chlorinity range of $0.00\~6.64\%_{\circ}$ (optimun chlorinity : $2.21{\%}_{\circ}$). The relationship between incubation period (Y in days) and water temperature(X in $^{\circ}C$) could be expressed as Y= 50.803-1.3555X. The eggs hatched $12{\~}13$ days after oviposition at $28.0{\~}28.6^{\circ}C$ 3. The pre-spawning moltings were appreciably different in the morphologic structure from those of common moltings. Breeding setae and dresses were formed on the thoracic regions, abdominal epimerae and the bases of the first to fourth pleopods in order to prepare and support oviposition, transfering and supporting eggs in egg-chambers up to hatching. These supplementary breeding organs were observed only at reproductive seasons.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.28-28
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• 2003

Viable interspecific hybrids between yellowtail flounder (Pleuronectes ferrugineus, Store.) and winter flounder (Pleuronectes americanus, Walbaum) were produced by artificial insemination of yellowtail flounder eggs with winter flounder sperm. However, mean fertilization rate, hatching success and early survival up to 3 weeks post hatch were significantly lower than those of parental pure cross controls (P<0.01). Overall, cytogenetic traits (karyological analysis and estimation of cellular DNA contents using flow cytometry) of hybrid flounder were intermediate between the two parental species. Microsatellite assay was used to distinguish the parental genomes in the hybrids; in most cases, one allele was specific to each of the parents. Morphometrics assessed by body proportions indicated that hybrids generally displayed a morphology intermediate between the maternal and paternal species. Interspecific hybrids exhibited abnormal and retarded gonad development in both sexes based on histological analysis of gonads from adult fish. The sterility of the hybrids presents a significant advantage for their use in aquaculture, as potential escapees would not be capable of reproducing in the wild and contaminating natural stocks.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.2
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• pp.196-200
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• 1998

The proximate composition of viscera of six fish species viz., Catla catla, Labeo, rohita Cirrhina mrigala, Hypophthalmichthys molitrix, Ctenopharyngodon idella and Cyprinus carpio under the influence of artificial feed $(T_1)$, broiler manure $(T_2)$, buffalo manure $(T_3)$, N:P:K (25:25:0 $T_4$) and control pond $(T_5)$ has been studied on final harvest after the end of experimental period of one year. The overall comparison of six fish species under five different treatments showed that Cyprinus carpio remained the best for visceral total protein contents. The maximum visceral protein content was recorded in fishes under $T_2$ except Cirrhina mrigala, while the minimum under $T_5$. The comparison of five treatments showed that maximum (6.69) percentage of fat in viscera of fish was recorded under $T_2$, closely followed by $T_4$ while minimum (4.89) under the effect of $T_5$. Among the six fish species under five treatments Cyprinus carpio showed maximum accumulation/deposition of total fats in viscera.

• The Journal of Korean Obstetrics and Gynecology
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• v.29 no.3
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• pp.68-77
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• 2016

Objectives: This paper is to report a spontaneous pregnancy with low AMH level and thin endometrium infertility after Korean medical treatments.Methods: A 38-year old patient who had low AMH level, thin endometrium, sleep disturbance and anxiety happened after anorectal cancer surgery was treated by korean medicine such as herb medications per day for 6 months. And we observed the effects of treatments by improvement of symptoms checking endometrium ultrasonography.Results: After treatments, Symptoms of anxiety and sleep disturbance were improved. The number of ovarian follicle and endometrium thickness were upgraded during artificial pregnancy trials. Finally the patient was a spontaneous pregnancy after failures of one intrauterine insemination and one in vitro fertilization trial.Conclusions: This case shows that the korean medicine has effective option for a patient with low AMH level and thin endometrium infertility. Further ultrasonography would be valid measurements to check the effectiveness of oriental treatments as well as symptoms.

• Development and Reproduction
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• v.7 no.1
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• pp.9-13
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• 2003

A series of experiments were conducted to compare the effects of various diluents in cold storage on the sea urchin, Hemicentrotus pulcherimus sperm. Various diluents of glucose solutions, artificial sea water(ASW) and 50% ASW were used to store the sperm at 4$^{\circ}C$. The storage effect was evaluated using sperm activity index(SAI), survival rate of sperm and fertilization rate to egg. ASW and 1.2 M glucose were found to be better diluents which maintained high motility and survival rate of sperm f3r a storage period of 30 days. Optimal pH of diluent to store the sperm at 4$^{\circ}C$ is 7.0∼8.0. In order to keep high SAI and survival rate of sperm, addition of 400 ppm neomycin into the diluent revealed the best storage results.

• Development and Reproduction
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• v.21 no.3
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• pp.229-235
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• 2017

In the field of reproductive medicine, assessment of sperm motility is a key factor for achieving successful artificial insemination, in vitro fertilization, or intracellular sperm injection. In this study, the motility of boar sperms was estimated using real-time imaging via confocal microscopy. To confirm this confocal imaging method, flagellar beats and whiplash-like movement angles were compared between fresh and low-temperature-preserved ($17^{\circ}C$ for 24 h) porcine sperms. Low-temperature preservation reduced the number of flagellar beats from $11.0{\pm}2.3beats/s$ (fresh sperm) to $5.7{\pm}1.8beats/s$ and increased the flagellar bending angle from $19.8^{\circ}{\pm}13.8^{\circ}$ (fresh) to $30.6^{\circ}{\pm}15.6^{\circ}$. These data suggest that sperm activity can be assessed using confocal microscopy. The observed motility patterns could be used to develop a sperm evaluation index and automated confocal microscopic sperm motility analysis techniques.

• Fisheries and Aquatic Sciences
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• v.4 no.1
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• pp.39-46
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• 2001

To examine the effect of copy number-dependent transgenic genotype on the expression of foreign gene, stable hemizygous and homozygous transgenic breeding line was established using artificial parthenogenesis. For this purpose, induced diploid gynogenetic transgenesis was optimized in mud loach (Misgurnus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to $4,050\;ergs/mm^2$ The UV-irradiated sperm were inseminated into eggs from recessive color strain (yellow) or heterozygous transgenic mud loach containing CAT gene. Cold shock at $2^{\circ}C$ for 60 min, 5 min post fertilization successfully restored the diploidy of eggs inseminated with UV-irradiated sperm. Restoration to diploidy was confirmed by flow cytometry and gynogenetic status was verified by examining maternal exclusive inheritance of multi-locus DNA fingerprints, body color and transgenic marker. Putative isogenic transgenic fish clearly showed homozygous status at trans gene locus based on Southern blot hybridization and progeny testing. Further, such homozygous gynogenetic diploids revealed the increased levels of transgene expression, when compared to those of heterozygous (hemizygous) transgenic fish.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.740-744
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• 2002

The relationship of gonad weight and liver weight with body weight of six fish species viz; Catla catla, Labeo rohita, Cirrhina mrigala, Hypophthlamichthys molitrix, Ctenopharyngodon idella and Cyprinus carpio under the influence of artificial feed, broiler manure, buffalo manure, N:P:K (25:25:0) and a control pond were examined after a rearing period of one year. The positive relationship between gonad weight and body weight was significant which showed the dependence of gonadal development on body weight in all the six fish species. The currelation coefficients were higher in female fishes. However, the major carps had a much smaller proportional gonadal weight as compared with Chinese carps and a common carp due to their faster growth rate. The overall comparison of six fish species under different experimental treatments revealed that highest liver weight was observed for C. idella closely followed by C. carpio. The maximum correlation value was observed with H. molitrix under the broiler manure. The maximum Gonadosomatic Index (GSI) remained as 32.63 for C. carpio followed by C. idella. The maximum value for Hepatosomatic Index (HSI). remained 1.99 for C. idella followed by C. mrigala.

• Journal of Embryo Transfer
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• v.11 no.1
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• pp.71-80
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• 1996

The envelope of the rnannnalian oocyte plays crucial roles in sperm-oocyte interactions by providing sperm receptors, inducing acrosome reaction and preventing polyspermy. Understanding of properties of the zona pellucida (ZP) is essential for the artificial control of fertility in mammals. This study was carried out to produce and characterize monoclonal antibodies(MAbs) to porcine ZP proteins. Approximately 8,000 ZPs were obtained from follicular oocytes and dissolved in 40$\mu$l of double distilled water. Following immunization through foot-pad injections of Balb /c mice with a ZP solution, the popliteal lymph nodes were recovered at 2 weeks after the last injection. Hybridoma cell lines were established by fusing lymph node cells with P3X63 myeloma cells through selection using HAT medium and screening by immunofluorescence(IF) microscopy on the isolated ZP. Secreted MAbs were found to consist k chains and different heavy chains as evidenced by isotyping. Some of the MAbs demonstrated high specificity to the ZP in IF. The Mabs also showed positive cross reactivity with hamster and mouse eggs, while negative with bovine eggs. The results implicate that the MAbs can be used not only for identification of functional regions of the ZP, but also for elucidation of mechanisms involved in fertilization of mammals. The MAbs will provide basic information on biochemical anatomy of the ZP as well as can be candidates for the future contraceptive vaccines.