• Title/Summary/Keyword: Fertilized egg development

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A Study on the Induction of Infertility of Largemouth Bass (Micropterus salmoides) by CRISPR/Cas9 System (CRISPR/Cas9 System을 활용한 배스의 불임 유도에 대한 연구)

  • Park, Seung-Chul;Kim, Jong Hyun;Lee, Yoon Jeong
    • Korean Journal of Environment and Ecology
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    • v.35 no.5
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    • pp.503-524
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    • 2021
  • A largemouth bass (Micropterus salmoides) is an ecosystem disturbance fish species at the highest rank in the aquatic ecosystem, causing a serious imbalance in freshwater ecosystems. Although various attempts have been made to eradicate and control largemouth bass, no effective measures were found. Therefore, it is necessary to find an approach to maximize the effective population reduction based on the unique characteristics of largemouth bass. This study used the transcriptome analysis to derive 182,887 unigene contigs and select 12 types of final target sequences for applying the CRISPR/Cas9 system in the genes of IZUMO1 and Zona pellucida sperm-binding protein, which are proteins involved in sperm-egg recognition. After synthesizing 12 types of sgRNA capable of recognizing each target sequence, 12 types of Cas9-sgRNA ribonucleoprotein (RNP) complexes to be used in subsequent studies were prepared. This study searched the protein-coding gene of sperm-egg through the Next Generation Sequencing (NGS) and edited genes through the CRISPR/Cas9 system to induce infertile individuals that produced reproductive cells but could not form fertilized eggs. Through such a series of processes, it successfully established a composition development process for largemouth bass. It is judged that this study contributed to securing the valuable basic data for follow-up studies to verify its effect for the management of ecological disturbances without affecting the habitat of other endemic species in the same water system with the largemouth bass.

Morphological of Development Eggs, Larvae and Juveniles Gymnogobius urotaenia in Hwangbocheon, Korea (황보천에 서식하는 꾹저구 Gymnogobius urotaenia의 난발생 및 자치어 형태발달)

  • Jae-Min Park;Kyeong-Ho Han
    • Korean Journal of Ichthyology
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    • v.35 no.4
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    • pp.253-262
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    • 2023
  • This study compared the results of observing the early life history of Gymnogobius urotaenia, which lives in Hwangbocheon Stream, an inflow stream on the east coast, with the differences between fish of the same Gobiidae fishes. In May 2022 and 2023, fertilized eggs and brood stork scattered under rocks were captured twice in Hwangbocheon Stream. The spawning amount was 827~1,540 and the orchid was a elliptical in shape with a size of 3.21×1.07 mm. The stage of ovulation observed in the laboratory was 16 cells, and hatching began after 193 hours. The breeding water temperature range was 18.8~19.3℃. Newly after hatching larvae, the yolk sac with a total length of 3.84~4.33 (average 4.10±0.17, n=30) mm, and the anus was not open. 6 days after hatching of incubation, the total length was 5.32~6.11 (average 5.67±0.25, n=30) mm, absorbing all egg yolk and transitioning to the preflexion larvae, ingesting food, and developing a keynote on the tail fin. 15 days after hatching, the end of the urostyle end was completely bent at 45° with a total length of 7.33~8.52 (average 7.81±0.46, n=30) mm and transitioned to postflexion larvae, and melanophore developed throughout the body. 38 days after hatching, the total length is 22.1~26.1 (23.8±1.36, n=30) mm and the number of fins (6 first dorsal fins, 11 second dorsal fins, and 11 anal fins, 12 ventral fins) is all the number of fin base became integer and transferred to the juvenile. As a result of the study, it was possible to distinguish the melanophore of postflexion larvae of G. urotaenia from other postflexion larvae Gobiidae fish in that they were distributed throughout the body of half the body and tail.

Egg Development and Morphological Change of Larvae and Juveniles of the Starry Flounder, Platichthys stellatus (강도다리, Platichthys stellatus의 난발생과 자치어의 형태발달)

  • Byun, Soon-Gyu;Lee, Bae-Ik;Lee, Jong-Ha;Ku, Hak-Dong;Park, Sang-Un;Yun, Seong-Min;Hwang, Seon-Young;Kim, Yi-Cheong;Han, Hyung-Gyun
    • Korean Journal of Ichthyology
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    • v.19 no.4
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    • pp.350-359
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    • 2007
  • The egg development and morphological change of larvae and juveniles of the starry flounder, Platichthys stellatus were observed in laboratory. Fertilized eggs of the species, 1.09~1.19 mm (mean $1.13{\pm}0.03mm$, n=50) in diameter, were floating, colorless, transparent in shape and lacked in oil globules. The eggs hatched out 121 hours after fertilization at water temperature $8.2{\sim}11.2^{\circ}C$. The size of the hatched larvae were 2.58~2.89 mm (mean $2.67{\pm}0.09mm$) in total length, their mouth and anus were not open yet and myotome number was 14+27=41. Melanophore and xanthophore appeared on the notochord and digestive organ and the margin of membrane fin, on the yolk sac and eyes were lacking in pigment cells. 5 days after hatching the larvae attained 4.30~4.97 mm (mean$4.74{\pm}0.21mm$) in TL, and their mouth and anus were open. 10 days after hatching the larvae transformed to postlarval stage and they were 4.67~5.75 mm in TL (mean $5.30{\pm}0.31mm$), and absorbing the yolk completely. Feeding activity increased as the mouth became larger. At 23 days, the larvae attained 6.69~8.82 mm in TL (mean $7.85{\pm}0.75mm$), and the right eye was started moving to the left side of the head. At 52 days, the juveniles attained 10.99~17.06 mm in TL (mean $13.50{\pm}1.67mm$). The right eye was moved completely onto the left side. All of the fins had completed set of the fin rays (D. 64~67: A. 45~51: P. 11: V. 6: C. 19).

Effects of Bisphenol A on the Hatching of fertilized Eggs and Spawning of Adult Fish in Songsari, Oryzias latipes (Bisphenol A가 송사리, Oryzias latipes의 수정란 부화와 어미의 산란에 미치는 영향)

  • NA Oh Soo;OH Seong Rip;LEE Young Don;BAEK Hea Ja;KIM Hyung Bae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.5
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    • pp.378-382
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    • 2000
  • The effects of bisphenol A (BPA), known as one of the endocrine disrupting chemicals (EDCs), on the hatching of fertilized eggs (FE) and the spawning of adult fish in songsari, Oryzias latipes were investigated. In condition of the experimental groups, incubated water temperature was $25.0{\pm}0.5^{\circ}C$ and photoperiod was 16 h (light) : 8 h (dark). fE were exposed to aqueous solutions of BPA at nominal concentration of 50, 100 and $200 {\mu}g/l$. The time required in hatching of FE was long in the BPA $50 and 100 {\mu}g/l$ treatment groups when compared to the controls, and in the BPA $200 {\mu}g/l$ treatment group, FE failed to hatch. And also hatching rate was decreased in the BPA treatment groups in comparison to the controls, Adult fishes were reared to oral administration via a diet of 50, 100 and $200 {\mu}g/g$ body weight BPA concentration for 3 weeks, Number of total eggs spawned in the adult fish were fewer in the BPA 100 and 200 treatment groups than in the controls and BPA 50 treatment group. Frequency of abnormal eggs in the total eggs spawned was 11.8 and $16.2{\%}$ in the control and acetone carrier control lower than 36.8, 46,8 and $74.1{\%}$ in the BPA 50, 100 and 200 treatment groups, respectively. And hatching rate of FE decreased in the BPA treatment groups in comparison to the controls. In these results, inhibition of the hatching of FE and the quantity and quality of spawned eggs in adult fish were observed from BPA treatment groups when compared to the controls.

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Development and Growth of Larvae of Four Bivalve Species (4종 조개류 유생의 발생과 성장)

  • 허영백;허성범
    • Journal of Aquaculture
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    • v.13 no.2
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    • pp.119-128
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    • 2000
  • Exposure to air and increased temperature induced successful spawning in Mytilus edulis, M. coruscus Crassostrea gigas and Pinctata fucata martensii. Developmental durations required for an egg to attain D-shaped larva and the D-shaped larva to reach pediveliger stage were estimated in these bivalves. Size of fertilized eggs was the largest (70.3 ${\mu}m$) in M. coruscus and the smallest (45.3 ${\mu}m$) in P. fucata martensii. At 17$^{\circ}C$, M. edulis and M. coruscus attained D-shaped larval stage within 48 hours after fertilization but those of C. gigas and B. fucata martensiii within 24 and 22 hours at 21 and 26$^{\circ}C$, respectively. The development duration required for a D-shaped larva to attain pediveliger stage was the longest (27 days) in M. coruscus and ranged between 20 and 22 days for the others. The shell length of the pediveliger was the longest (274.9 ${\mu}m$) in C. gigas and smallest (190.9 ${\mu}m$) in P. fucata martensii Length and height of larval shell was highly correlated with each other in all the 4 species. The shell height of C. gigas was more than the shell length beyond the size of 100 ${\mu}m$ shell length. However, shell length of the others was always longer than shell height at the larval stage.

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STUDIES ON THE PROPAGATION OF TOP SHELL-I Spawning and early development of the top shell, Turbo cornutus SOLANDER (소라 Turbo cornutus SOLANDER의 증식에 관한 연구-I 소라의 산란과 초기발생)

  • RHO Sum
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.9 no.1
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    • pp.43-55
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    • 1976
  • Fertilization and early development of turbo cornutus was studied based on the samples which were collected in Yeosu area. Particular emphasis was paid on induction of artificial spawing, fertilization rate, preembryonic development, the growth of the early larva and larval survival to various salinity. Among the various methods for induction of artificial spawning which have been tested for the present study, drying by exposure to air is the. most efficient, and percentage fertilization rate was $83.8-96.4\%$. The diameter of fertilized eggs was $0.182{\pm}0.0028mm$; and the diameter of egg membrane was $0.245{\pm}0.093mm$. Under the temperature range of $20.6-25.4^{\circ}C$ the larvae hatched out after 11:05-11:15 hours of fertilization. After 3.0-3.5 days of fertilization the planktonic larvae begand to settle, and the settlement terminated within 5 days. During the period of 150 days of early culturing the diameter growth of shell(M) and the diameter of shell aperture(A) was formulated as follows: $$1972\;M=0.33e^{0.02070D}$$ $$A=0.19e^{0.02282D}$$ $$1973\;M=0.32e^{0.02282D}$$ $$A=0.16e^{0.02596D}$$ During the same period of early culturing the relative growth of shell diameter and the diameter of shell aperture was formulated as follows : 1972 A=0.6478 S-0.1575 1973 A=0.5897 S-0.0515 After 11 days of larval hatching $0.02-0.18\%$ of planktonic larvae settled. After 150 days of settlement the survival rate of the early shells was $7.4-21.6\%$. Under the temperature range of $21.0-22.7^{\circ}C$ the optimum salinity range for the development of egg and the planktonic larvae was $30-35\%_{\circ}$.

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Breeding and Reproductive Studies on Korean Native Loach III. Early Embryonic Development (한국산 미꾸리에 관한 육종 번식학적 연구 III. 초기 난발생)

  • 이종영;윤종만;이재현;신재구;박홍양
    • Korean Journal of Animal Reproduction
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    • v.16 no.1
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    • pp.63-76
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    • 1992
  • This study was carried out to investigate the appearance time of the second polar body for producing Gynogenesis or Triploid which could be obtained by arresting the second polar body by cold shock, and then blastoderm was used to measure fertility that revealed the nature of oogenesis, the effects of water temperature on fertility, hatchability, abnormality, viability and growth rate, and the water temperature and the breeding methods to prevent early death of larvae in Korean loach(Misgurnus anguillicaudatus) ; the results obtained in this study were summarized as follows. The second polar body was observed ont he surface of plasma disc close to micropyle within 10~40 min after fertilization at 29$^{\circ}C$. Artificial inseminatin had to be done immediately after the egg spawning because the spermatozoa of loach their mobility within 2 minutes when they were exposed to water. The amount of time needed to reach at blastoderm stage was 12 hours if fertilized eggs were incubated at 16$^{\circ}C$, 8 hours at 19$^{\circ}C$, 6 hours at 21$^{\circ}C$, 5 hours at 23$^{\circ}C$, 4 hours at 26$^{\circ}C$ and 3 hours 30 min at 29$^{\circ}C$ showing the shorter time for development of eggs at higher temperature. Fertilization rates in water temperatures of 19$^{\circ}C$, 21$^{\circ}C$, 23$^{\circ}C$, and 26$^{\circ}C$ were higher than those of water temperatures, 16$^{\circ}C$ and 29$^{\circ}C$. Water temperatures at 19$^{\circ}C$, 21$^{\circ}C$, and 23$^{\circ}C$ showed higher hatching rates that those of 16$^{\circ}C$, 26$^{\circ}C$, and 29$^{\circ}C$, while abnormal rates in 16$^{\circ}C$, 19$^{\circ}C$, 21$^{\circ}C$ and 23$^{\circ}C$ were lower than that of 26$^{\circ}C$ and 29$^{\circ}C$. Water temperatures at 16$^{\circ}C$, 19$^{\circ}C$, 21$^{\circ}C$, 23$^{\circ}C$ and 26$^{\circ}C$ respectively, were more different than 29$^{\circ}C$ in survival rates. The embryos were hatched at 72 hours after fertilization in 16$^{\circ}C$ water temperature, 48 hours in 19$^{\circ}C$, 40 hours in 21$^{\circ}C$, 32 hours in 23$^{\circ}C$, 25 hours in 26$^{\circ}C$, and 16 hours in 29$^{\circ}C$. Within three days after hatched out, the larvage grew 3mm in total length, the yolk granules were entirely consumed and the head and the trunk became thicker. Within 45 days after hatched out, the larva grew 25mm at 29$^{\circ}C$, 21mm at 26$^{\circ}C$, 16mm at 23$^{\circ}C$, 15mm at 21$^{\circ}C$, 12mm at 16$^{\circ}C$ in a 30 litreglass aquarium.

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Spawning Character and Early Life History of the Endangered Korean Dwarf Loach, Kichulchoia brevifasciata (Teleostei: Cobitidae) (멸종위기종 좀수수치 Kichulchoia brevifasciata의 산란기 특징 및 초기생활사)

  • Ko, Myeong-Hun;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.26 no.2
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    • pp.89-98
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    • 2014
  • Eggs development and early life history of the endangered Korean dwarf loach, Kichulchoia brevifasciata (Teleostei: Cobitidae) was investigated to provide basic information regarding biological characteristics and restoration. Adult fish specimens were sampled using a spoon net at Geurnsan-myeon, Goheung-gun, Jeollanam-do, Korea from June to July 2011. Since, spawning characteristics were analyzed, and females were induced to spawn by injecting Ovaprim (0.5 mL/kg) and their eggs were artificially fertilized with sperms by the dry method in the laboratory. Total length of mature female were 46~76 mm with GSI $9.6{\pm}3.77%$, and total length of mature male was 42~52 mm with GSI $3.5{\pm}1.04%$. Sex ratio (♂/♀) was 0.10, and there were no secondary sexual characteristics. The number of mature eggs was averaged $60{\pm}28.7$ per female. The lemon yellow eggs were slightly adhesive $1.46{\pm}0.07mm$ in diameter. The embryo hatched approximately 66 h after fertilization at $25^{\circ}C$, and the hatched larvae were averaged $5.5{\pm}0.07mm$ in total length (TL). At 6 days after hatching, the larvae averaged $9.0{\pm}0.29mm$(TL) and their yolk sac was completely absorbed. At 17 days after hatching, they entered the juvenile stage and reached $12.6{\pm}0.24mm$ (TL). At 80 days after hatching, the band patterns and external form of the juveniles were similar to those of adults, and they averaged $33.0{\pm}2.19mm$(TL).

Embryonic and Larval Development of Slender Catfish, Silurus microdorsalis Mori, 1936, Endemic to Korea (한국 고유종 미유기(Silurus microdorsalis Mori, 1936)의 난 발생 및 자치어 형태발달)

  • Kang-Rae Kim;Yeong-Ho Kwak;Mu-Sung Sung;Heon Yang;Seong-Jang Cho;Bong Han Yun;In-Chul Bang
    • Korean Journal of Ichthyology
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    • v.35 no.2
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    • pp.91-100
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    • 2023
  • The early life history of Silurus microdorsalis living in Jahocheon Stream was studied by observing egg and morphological development. Live fish were captured in June 2018, then reared in a circulating filtration system under a 14L : 10D photoperiod with a water temperature of 18℃. To artificially induce spawning, females were injected with 0.5 mL of Ovaprim (Syndel, Nanaimo, BC, Canada) per kg of body weight, and males were injected with 10,000 IU/kg body weight of human chorionic gonadotropin. Approximately 15 h later, eggs were artificially inseminated by the dry method. Mature eggs were light pale yellow, which separated them from immature eggs. Fertilized eggs were 2.16±0.06 mm (n=8) in diameter and fully hatched at 181 h after fertilization. The fertilization rate was 63.1±2.2%, and 10.0±3.7% of the embryos were malformed at 18℃. The rates of development were 181 h at 18℃, 109 h at 21℃, and 76 h at 24℃. The larval size immediately after hatching was 4.64±0.22 mm (n=8), and the larvae displayed negative phototaxis at 1 day after hatching. The total larval length on 7 days after hatching was 12.47±0.53 mm, with 25~30 basal anal fin rays and 14~16 basal caudal fin rays observed. The total larval length was 14.13±0.51 mm on 9 days after hatching, and approximately 90% of the black endoplasmic reticulum was deposited on the head and body. The dorsal fin had formed, and a single basal body was observed. On 15 days after hatching, the total larval length was 16.69±0.31 mm; the number of basal caudal fin rays (18 poles) was an integer because 2 dorsal fin basal rays and 60~63 anal fin basal rays were observed. The total larval length was 28.96±1.10 mm on 50 days after hatching; the numbers of caudal fins (n=18), dorsal fins (n=3), pectoral fins (n=11), and anal fin basal rays (n=67~73) were integers.

Studies on the Bionomic of Gonatocerus sp. y (Hymenoptera: Mymaridae), an Egg Parasite of the Green Rice Leafhopper, Nephotettix cincticeps Uhler (끝동매미충 난기생봉(Gonatocerus sp. y)의 생태에 관한 연구)

  • Kim Jeoung Bu;Kim Chang Hyo
    • Korean journal of applied entomology
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    • v.23 no.3 s.60
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    • pp.158-165
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    • 1984
  • These studies were carried out to investigate the bionomics of Gonatocerus sp. y (Hymenoptera: Mymaridae) , an egg parasite of the green rice leafhopper, Nephotettix cincticeps Uhler (Homoptera: Deltocephalidae) under laboratory and field conditions at Gyeongnam Provincial O.R.D. in Jinju from 1982 to 1983. The developmental period of each life stage was observed under four different temperature conditions from $15^{\circ}C\;to\;30^{\circ}C\;with\;5^{\circ}C$ increment. The development from egg to adult at 15^{\circ}C,\;20^{\circ}C,\;25^{\circ}C,\;and\;30^{\circ}C $ was completed in an average of 53, 25, 15, and 12 days and the females laid an average of 28, 48, 42, and 32 eggs respectively, with a maximum of 98. And most of the eggs were laid in the first two days after emergence. The time of adult emergence was determined under $25{\pm}1^{\circ}C$ condition. More than $76\%$ of the wasps emerged in the morning and the rest of them emerged in the afternoon. The average sex ratio between male and female was 1 : 5 when the wasps were developed from fertilized eggs. The longevity of female was greater than that of male when honey and water were provided as food, however, it was not when only water was supplied as food. The average longevities of females were 30 days at $15^{\circ}C$ and 15 days at $20^{\circ}C$ when honey and water were given as food. The peak precent parasitism of Gonatocerus sp. v on Nephotettix cincticeps in fallow fields was observed between late April and early May. The percent parasitism of the wasps In fallow fields was an average of 49\~59.9\% $ in Gyeochang, Namji and $23.2\~36\%$ in Namhae. The parasitic activities of the parasite had four peaks in paddy field, however, the highest peak was observed between late August and early September. The Percent e99 Parasitism of Gonatocerus sp. y was an average of $12.3\%$ on second the generation and $24.6\%$ of third genera lion of Nephotettix cinrticeps.

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