• Title/Summary/Keyword: fertilized eggs

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Studies on In Vitro Fertilization and Development of In Vitro Matured Porcine Follicular Oocytes I. Effect of Various Media and Co-culture with Porcine Cumulus Cellsor Mouse Fetal Fibroblast Cells on In Vitro Development of In Vitro Fertilized Oocytes (체외성숙 돼지난포란의 체외수정과 배발달에 관한 연구 II. 각종 배양액, 돼지난구세포 및 생쥐태아간세포와의 공동배양이 체외수정 돼지 난포란의 체외발달에 미치는 영향)

  • 정형민;엄상준;승경록;이상준;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.113-120
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    • 1993
  • To provide the optimal culture conditions for the developm,ent of in vit개 produced embryos, we have been investigated various culture media as well as co-cultrue systems using porcine cumulus cells or mouse fetal fibroblast cells. Porcine ovaries were brought to the laboratory from local slaughter house within 1 hour after slaughtering and cumulus oocytes complexes were recovered from antral follicles(3~5mm) with 23 gauge needle. To maturate follicular oocytes, cumulus oocytes complexes were washed three times with TCM-199 containing 25mM HEPES and incubated(39$^{\circ}C$, 5% CO2 in air) in various maturation media for 42 hrs. Ejaculated and liquid storaged boar spermatozoa capacitated with different sperm capacitation methods and media were rpepared for fertilizing of matured follicular oocytes in vitro. Fertilization was performed by adding 5~10${\mu}\ell$ fo capacitated spermatozoa containing 1~5$\times$105 sperm/ml to droplets. Eighteen to twenty-eight hours after sperm insemination, fertilized eggs were washed three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three times with culture media and transferred to the various culture media, to the culture media with a monolayer of somatic cells. The in vitro development rates of 1-cell embryos cultured with three different media, m-KRB, BECM and TCM-HEPES were 0~1.0%, showing extremely lower rates. Especially, most of embryos were observed to arrest the development beyond 4-cell stages. The rates of embryos developed to 2-, 4-, 8-, 16-, 32-cell and morula or blastocyst stage in co-culture with porcine cumulus cells and mouse fetal fibroblast cells were 61.1~67.0%, 59.0~58.0%, 42.5~43.1%, 28.4~30.2% and 20.4~21.0%, respectively. These development rates upto morula or blastocyst stages were significantly higher than those of the embryos cultured in the basic culture medium(P<0.01). These findings suggest that co-culture of in vitro fertilized eggs with porcine cumulus cells or mouse fetal fibroblast cells enhance the development of fertilized eggs to morula or blastocyst stage in vitro.

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Morphological Development of Embryo, Larvae and Juvenile in Yellowtail kingfish, Seriola lalandi

  • Yang, Sang Geun;Hur, Sang Woo;Ji, Seung Cheol;Lim, Sang Gu;Kim, Bong Seok;Jeong, Minhwan;Lee, Chi Hoon;Lee, Young-Don
    • Development and Reproduction
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    • v.20 no.2
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    • pp.109-118
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    • 2016
  • This study monitored the morphological development of embryo, larvae and juvenile yellowtail kingfish, Seriola lalandi, for their aquaculture. The fertilized eggs obtained by natural spawning were spherical shape and buoyant. Fertilized eggs were transparent and had one oil globule in the yolk, with an egg diameter of $1.35{\pm}0.04mm$ and an oil globule diameter of $0.32{\pm}0.02mm$. The fertilized eggs hatched 67-75 h after fertilization in water at $20{\pm}0.5^{\circ}C$. The total length (TL) of the hatched larvae was $3.62{\pm}0.16mm$. During hatching, the larvae, with their mouth and anus not yet opened. The yolk was completely absorbed 3 days after hatching (DAH), while the TL of post-larvae was $4.72{\pm}0.07mm$. At 40 DAH, the juveniles had grown to $30.44{\pm}4.07mm$ in TL, body depth increased, the body color changed to a black, yellow, and light gray-blue color, and 3-4 vertical stripes appeared. At 45 DAH, the juveniles were $38.67{\pm}5.65mm$ in TL and $10.10{\pm}0.94mm$ in body depth. The fish were green with a light orange color, with 7 faint green-brown stripes on the sides of their body. At 87 DAH, the juveniles had grown to 236.11 mm in TL, 217.68 mm in fork length, and 136.5 g in weight. The fish resembled their adult form, with a light yellow-green body color, loss of the pattern on the sides of their body, and a yellow coloration at the tip of the caudal fin.

Cross Fertility between the Wild Japanese Quail in the Weishan Lake Area and Domestic Quail

  • Xu, W.;Chang, H.;Wang, H.Y.;Chang, G.B.;Du, L.;Lu, S.X.;Yi, H.Q.;Xu, Q.;Xu, M.;Wang, Q.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.10
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    • pp.1421-1423
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    • 2003
  • Cross fertility between wild Japanese quail and domestic quail was explored in an experiment conducted on 18(3♂, 15♀)wild Japanese quails in Weishan Lake area, 18(3♂, 15♀)medium-sized domestic quails and 18(3♂, 15♀)pint-sized domestic quails, which were divided into nine groups. This study demonstrated that wild quail could succeed in crossing with domestic quail,producing fertilized eggs and hatching first filial generation. The findings indicated that there were no reproduction isolation between the wild Japanese quail and domestic quail, and that the best cross combination was between wild male quail and medium-sized domestic female quail, in which the fertility rate and hatchability of fertilized eggs amounted to 42.86% and 29.63% respectively. Based on the results, a new way could be adopted to protect, exploit and utilize genetic resources of wild quail.

Effects of pH on Fertilization and the Hatching Rates of Far Eastern Catfish Silurus asotus

  • Gao, Yang;Kim, Sun-Gyu;Lee, Jeong-Yeol
    • Fisheries and Aquatic Sciences
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    • v.14 no.4
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    • pp.417-420
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    • 2011
  • In this experiment, we examined the survival, fertilization, hatching times, and hatching rates of Far Eastern catfish Silurus asotus at pH ranging from 2 to 13 under laboratory conditions. Eggs could be fertilized at pH 3-12. In a hatching experiment, mortality was first observed at pH 13, when all fertilized eggs died within 8 min, followed by pH 2 (30 min), pH 12 (60 min), pH 3 (4 h), and pH 11 (5 h). Hatching only occurred at pH 4-10, with the highest hatching rate at pH 7 (52%) and the lowest at pH 10 (24%). Hatching rates in acid solutions were higher than in alkaline solutions, although the difference was not significant. Hatching was first observed at pH 10, beginning 27 h after fertilization and ending at the 31 h. A clear difference was observed between hatching times, ranging from 31 to 64 h and increasing in order with descending pH.

A Microscopic Study on the Egg Envelope of an Endemic Korean Fish, Coreoleuciscus splendidus, Cyprinidae, Teleostei

  • Kim, Chi-Hong;Park, Jong-Sung;Kim, Jae-Goo;Park, Jong-Young
    • Applied Microscopy
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    • v.44 no.3
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    • pp.96-99
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    • 2014
  • Study on the egg envelope of an endemic Korean freshwater fish, Coreoleuciscus splendidus was carried out by light and scanning electron microscopes during its spawning season. The egg envelope on the entire egg is filled with plenty of long cylinder-like villi before and after fertilization. The fertilized eggs consist of two regions, the attaching and non-attaching parts to stick to the substrates. The villi covering the fertilized eggs have three kinds of villi in its length: 1) normal- sized villi of an average of $10.6{\mu}m$ on the non-attachment part; 2) longer and more slender villi over about $20{\mu}m$ on the attachment part, at least being two times longer than those of the non-attachment; and 3) shorter villi under about $5.0{\mu}m$ around the micropyle, with half the size of the non-attachment villi. The micropyle rotated in a counter-clockwise direction with five pieces of the ridges.

Early Life Stage Toxicity of 2,3,7,8,-Tetrachlorodibenzop-Dioxin(TCDD) in Goldfish(Carassius auratus) (금붕어 수정난의 2,3,7,8-TCDD 노출에 따른 다이옥신 관련 유전자의 발현 및 형태학적 변화)

  • Oh, Seung-Min;Ryu, Byung-Taek;Kim, Ha-Ryong;Chung, Kyu-Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.24 no.1
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    • pp.1-8
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    • 2009
  • In this study, we obtained the fertilized eggs from goldfish(Carassius auratus) and observed normal developmental stage(from fertilized eggs to larvae) in non-exposed groups. Goldfish embryos at 3 h postfertilization(hpf) were statically exposed for 1 h to either dimethylsufoxide(DMSO, 0.1%, v/v) or TCDD($0.5{\mu}g/L$). Toxicity and morphological changes were characterized from 3 to 120 h postfertilization(hpf). Egg mortality($0{\sim}48$ hpf) and hatching ratio($72{\sim}83$ hpf) in TCDD-exposed group were significantly different from control groups. However, pericardial edema was first observed at 72 hpf, followed by the onset of yolk sac edema and mortality. In addition, goldfish embryos-larvae exposed to TCDD significantly increased TCDD-related gene such as CYP1A($24{\sim}72$ hpf) and AhR2(72 hpf). This is the first study about in-depth characterization of TCDD-induced developmental toxicity in goldfish(Carassius auratus).

Studies on the form and character of silkworm, Bombyx mori L. which bred from double copulation (동품종 교배와 이품종 교배를 교번한 이중교배의 차대잠 형질에 관한 연구 (1))

  • 김윤식
    • Journal of Sericultural and Entomological Science
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    • v.5
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    • pp.19-23
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    • 1965
  • Author studied to investigate the form and character of silkworm, Bombyx mori L. which bred from double copulation. The results obtained are as follows. 1. Some eggs of silkworm were fertilized evenly 10 minutes copulation, but it was varried by the race of the worm. In silkworm eggs production, the male and female should be separated from the early stage of 5th instar in order to do not make non hybrid eggs. 2. It was very difficult to find the tendency of fertilization ratio between homo race copulation and hetero race copulation in crossing with mates of two different race for a female (double crossing).

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Development of Allotriploid Embryos from Female Olive Flounder Paralichthys olivaceus Crossed with Male Starry Flounder Platichthys stellatus (넙치(Paralichthys olivaceus)와 강도다리(Platichthys stellatus)간 유도된 잡종 3배체의 난발생)

  • Jung, Hyo Sun;Ko, Min Gyun;Lee, Hyo Bin;Kim, Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.628-634
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    • 2016
  • We investigated the characteristics and rate of development of allotriploid embryos derived from a cross between female olive flounder Paralichthys olivaceus and male starry flounder Platichthys stellatus. The allotriploidy was induced by cold shocking fertilized eggs three minutes post-fertilization at 3°C for 45 minutes. The average cellular DNA content of the allotriploid embryos was 2.06±0.03 pg/cell, which is equal to the sum of the cellular DNA content of a diploid olive flounder (1.42 pg/cell) and a haploid starry flounder (0.66 pg/haploid cell). The first cleavage, midblastula, gastrula and Kupffer's vesicle appearance stages of the allotriploid eggs began at 1.5, 8, 13 and 26 hours after cold shocking at 18°C, respectively. The developmental rate of allotriploid eggs was equivalent to that of diploid and triploid olive flounder eggs at 10, 14 and 18°C. However, the hatching times of allotriploid eggs, 7 h at 10°C, 5 h at 14°C and 4 h at 18°C, were earlier than those of diploid and triploid olive flounder.

Changes in the Specific Gravity of Pacific Cod Gadus macrocephalus, During the Early Life Stages (대구(Gadus macrocephalus)의 초기 발생시기의 비중변화)

  • Lee, Hwa Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.3
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    • pp.332-337
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    • 2018
  • The Pacific cod Gadus macrocepahlus, lays demersal eggs and the hatching larvae rise toward the surface layer of the ocean to feed. The change in the specific gravity of eggs and larvae was investigated to examine their vertical distribution and movement in the water column. The specific gravities of fertilized eggs and various size classes of larvae were measured using a density gradient apparatus. In total, the instantaneous specific gravity of 146 eggs and 225 larvae were measured. To prevent any disturbance in the gradient water column due to larval movement, 0.004% MS222 was used for anesthesia. Due to their high specific gravity, eggs spawned were deposited over the sea-bed of the spawning ground. The specific gravity of hatching larvae decreased abruptly. However, Pacific cod larvae still had a comparatively high specific gravity at hatching ($1.03655{\pm}0.00146g/cm3$, n=4, mean SL=3.62 mm) and their specific gravities tended to decrease as they grew. The specific gravity stabilized 6 days after hatching ($1.02590{\pm}0.00212g/cm3$, n=15, mean SL=4.67 mm) and the cod larvae were eventually able to float in the water column.

Egg Development and Morphology of Larva and Juvenile of the Chub Mackerel, Scomber japonicus (고등어(Scomber japonicus)의 난발생 및 자치어 형태발달)

  • Park, Chung-Kug;Yeon, In-Ho;Choi, Nak-Hyun;Heo, Seung-Joon;Han, Kyeong-Ho;Lee, Won-Kyo
    • Development and Reproduction
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    • v.12 no.3
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    • pp.207-213
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    • 2008
  • In order to monitor the developmental features of embryos, larvae, and juveniles of chub mackerel, Scomber japonicus, the fertilized eggs were obtained by artificial insemination. The fertilized eggs were spherical in shape and floated separately. Their membrane and yolk having 1 oil globule were transparent. The transparent eggs, having one oil drop in the yolk, were $0.94{\sim}1.02\;mm$ (mean, $0.95{\pm}0.03\;mm$) in diameter. The fertilized eggs started hatching at 51 hrs after fertilization in $20{\pm}0.5^{\circ}C$ water. The total length of the hatched larvae was $2.52{\sim}3.0\;mm$ (mean, $2.75{\pm}0.04\;mm$). At hatching, the larva, with the mouth and anus not opened yet, had yolk sack, $28{\sim}31$ myomeres and eyes with melanophore. Yolk completely 2 days after hatching and the total length of post-larvae $3.12{\sim}3.63\;mm$ (mean, $3.39{\pm}0.05\;mm$). At the 18 days after hatching, the tip of tail became curved at the end and the stems for pectoral, dorsal, and caudal fins appeared. Juveniles, having all firm rays, was $44.12{\sim}58.72\;mm$ (mean $55.95{\pm}6.74\;mm$) in TL 25 days after hatching.

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