• Development and Reproduction
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• v.20 no.3
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• pp.187-196
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• 2016

The objective of the current study was to determine acute plasma stress responses in two size groups of juvenile Epinephelus akaara (average body weight: $8.4{\pm}2.1$ and $3.3{\pm}0.6g$; 150 and 120 days after hatch, respectively) exposed to abrupt salinity drops (from 34 practical salinity unit, PSU seawater to 18, 10 PSU (experiment 1) or 26, 18, 10 PSU (experiment 2), respectively). Plasma glucose, glutamic oxalate transaminase, glutamic pyruvate transaminase, red blood cell counts, and gill histology were determined during 72 h exposure. Significantly increased plasma glucose, glutamic oxalate transaminase levels, and red blood cell counts were observed in fish exposed to 18 or 10 PSU. Histological changes, such as hyperplasia and lifting of epithelium in the gill secondary lamellae, were also observed in fish exposed to 18 or 10 PSU at 72 h post-drop. E. akaara exposed to sudden salinity drops to 18 or 10 PSU still seems to undergo the primary adjustment phase before fish reaches a new homeostasis, whereas fish exposed to 26 PSU seems to mount osmotic changes. Therefore, the no observed adverse effect levels for 72 h acute salinity challenge was 26 PSU in our study, and salinity drop to 18 PSU and below can possibly cause acute adverse effect, in which fish could be vulnerable to additional stresses such as a temperature changes or handling stress.

• Development and Reproduction
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• v.24 no.3
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• pp.215-224
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• 2020

Seawater adaptability of steelhead trout increases along with the increase in the size of the fish, independent of parr-smolt transformation. Three 96 h seawater challenge tests were conducted to determine the size at which seawater adaptability of steelhead trout develops. Plasma Na⁺ and Cl^- levels, moisture content, gill Na⁺/K⁺ ATPase activity, and mortality during the 96 h after direct transfer to seawater (32 ppt) were determined. Plasma Na⁺ and Cl^- levels in 50 g fish continuously increased during the 96 h after the transfer to seawater (p<0.05), but the levels in 100 and 150 g fish leveled off after 24 h (p<0.05). Both 100 and 150 g size steelhead trout maintained muscle moisture content (%) better than 50 g size fish (p<0.05). Gill Na⁺/K⁺ ATPase activity in the 100 g size group increased in a time-dependent manner after transfer to seawater (p<0.05), whereas activity in the 50 and 150 g sizes did not increase (p>0.05), for which a possible explanation was discussed. A mere 2.6% mortality in both the 50 and 150 g size groups was observed. In conclusion, the current results indicate that 50 g size steelhead trout did not show development of a high level of hypoosmoregulatory capacity, whereas fish in the 100 and 150 g size groups showed a high level in our experimental conditions. Therefore, the steelhead trout larger than a 100 g size is recommended for transfer to seawater culture.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.363-376
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• 1993

This study was taken to examine serum components concentrations and electrophoretic patterns of female tilapia(Oreochromis niloticus) living in 0$\textperthousand$, 10$\textperthousand$, 20$\textperthousand$, and 30$\textperthousand$ salt concentrations, respectively. The results obtained in these experiments were summarized as follows. The level of albumin and total protein showed changes in each salinity, but didn't significantly(P<0.05) change in Oreochromis niloticus. The level of BUN didn't significantly(P<0.05) change. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$, each calcium level in every salinity groups showed less than that of control, and didn't significantly change in 10$\textperthousand$, 20$\textperthousand$, 30$\textperthousand$ salinity. The level of calcium didn't significantly(P<0.05) change in each salinity. In 20$\textperthousand$ salinity, the level of cholesterol was at the highest peak. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$, each glucose level gradually decreased. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$, each glucose level gradually decreased. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$. In 30$\textperthousand$ salinity, the level of alkaline phosphatase was at the highest peak. The level of serum enzyme such as SGOT and SGPT was higher in seawater-adapted group than in freshwater group. The level of phosphorus chnage significantly(P<0.05) in each salinity. Correlation coefficient between serum albumin and glucose in 0$\textperthousand$ was ＋0.924. Correlation coefficient between serum SGOT and SGPT of individuals in 0$\textperthousand$ was ＋0.917. Fraction 1 of transferrin patterns of tilapia(Oreochromis niloticus) adapted in seawater was much thicker than that of transferrin patterns of individuals adapted in freshwater. Also fraction No. a wasn't observed in some individuals adapted in freshwater. These results showed that transferrin adapted in seawater relatively increased. Slight differences, that is, showed to be observed in total iron binding capacityand iron saturatin rate between tilapia adapted in freshwater and in seawater. The increase in total iron binding capacity was attributed to a rise in transferrin pressent in the first fraction of serum protein adapted in seawater. Accordingly, the serum iron levles seemed to be related to salinity($\textperthousand$).

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2002.11a
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• pp.141-144
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• 2002

Environmental estrogens are natural or synthetic substances present in the aquatic environment, especially in effluent from sewage treatment. However, the adverse effects of these estrogenic substances on fish reproduction are unknown. Di-2-ethylhexyl phthalate (DEHP) is the most common phthalate, which Ps used as a plasticizer in polyvinylchloride (PVC), and it is widespread in the environment and has been found in aquatic organisms and sediments. Therefore, juvenile common carps (Cyprinus carpio) were exposed to nominal concentrations of 17$\beta$-estradiol (E2) (0.5, 5, 50 $\mu\textrm{g}$/L) and DEHP (10, 100, 500 $\mu\textrm{g}$/L) for 21 days, to determine the adverse reproductive effects of these compounds on plasma vitellogenin (VTG) induction, sex steroid level, and gonad weight. Electrophoresis (SDS-PAGE) revealed that much of VTG was induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Enzyme-linked immunosorbent assay (ELISA) revealed that VTG was significantly induced in fish exposed to 5 and 50 E$_2$ $\mu\textrm{g}$/L, and combination of 50 E$_2$ $\mu\textrm{g}$/L with 10 and 500 DEHP $\mu\textrm{g}$/L, but none of DEHP exposure showed induction. Analysis of sex steroid levels in some fish revealed that testosterone (T) was detected in both male and female fish of the control and DEHP exposures, but none of fish exposed to 22 concentrations had detectable testosterone level. On the other hand, E$_2$ exposure induced 17$\beta$-estradiol in plasma of male fish, but there was no induction of 17$\beta$-estradiol in plasma of male fish exposed to DEHP. Comparison of gonadosomatic index (GSI) revealed that maximal E$_2$ exposure inhibited ovarian growth, but maximal DEHP exposure stimulated testicular growth. The results indicated that those comparisons can be a useful bio-indicator for determining adverse reproductive effect of endocrine disrupting chemicals (EDCs).

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.19-30
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• 2024

Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

• Development and Reproduction
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• v.20 no.3
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• pp.255-266
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• 2016

The optimum concentrations of clove oil as an anesthetic for olive flounder (Paralichthys olivaceus) and the stress response of the fish to clove oil anesthesia were determined over a range of water temperatures, and investigated in a simulated transport experiment using analysis of various water and physiological parameters. While the time for induction of anesthesia decreased significantly as both the concentration of clove oil and water temperature increased, the recovery time increased significantly (P<0.05). The plasma cortisol concentration in fish at each temperature increased significantly up to 12 h following exposure (P<0.05), then decreased to 48 h (P<0.05). The DO dissolved oxygen concentrations, pH values, and the fish respiratory frequencies decreased over 6 h following exposure to clove oil in all experimental groups (P<0.05), whereas the $NH_4{^+}$ and $CO_2$ concentrations in all experimental groups increased up to 6 h (P<0.05). The pH values and DO concentrations increased with increasing clove oil concentration (P<0.05) in the 6 h following exposure, and the $CO_2$ and $NH_4{^+}$ concentrations and the respiratory frequencies decreased with increasing clove oil concentration (P<0.05). The results of this experiment suggest that clove oil reduced the metabolic activity of olive flounder, thus reducing $NH_4{^+}$ excretion and $O_2$ consumption. In conclusion, clove oil appears to be a cost-effective and efficient anesthetic that is safe for use and non-toxic to the fish and users. Its use provides the potential for improved transportation of olive flounder.

• Development and Reproduction
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• v.21 no.3
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• pp.287-295
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• 2017

This study was conducted to observe egg and larvae morphological development of carp to obtain basic data for resource conservation and taxonomic research. Brood carp used in the research (total length 67.3-75.5 cm, average $71.0{\pm}3.45cm$) were bred in a circular rearing aquarium ($600{\times}300{\times}100cm$) using a running water system from January to July, 2015. Breeding water temperature was maintained at $23.0-25.0^{\circ}C$(average $24.0^{\circ}C$). Fertilized carp eggs were translucent and globular, and their size was 1.75-1.89 mm (average $1.82{\pm}0.06mm$). Blastoderms formed 10 min after fertilization and reached the two-cell stage 30 min after fertilization. Then, the embryo turned dark and exhibited melanophores, and blood started flowing from the heart across the egg yolk at 42 hrs and 50 min after fertilization. Hatching began 70 hrs and 26 min after fertilization larvae emerged through the egg membrane, starting from the head. The length of prelarvae immediately after hatching was 5.23-5.38 mm (average $5.31{\pm}0.11mm$) the mouth and anus were closed, and the pectoral fin was formed. Postlarvae at 18 days after hatching had a total length of 11.9-13.9 mm (average $12.9{\pm}1.40mm$), separate anal fin and back membranes, and fin ray. Juveniles fish at 35 days after hatching had a total length of 29.9-30.2 mm (average $30.1{\pm}0.13mm$), with the body covered with scales, and the same number of fin rays, color, and shape as their broodstork.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 1998.07a
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• pp.12-18
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• 1998

Tremendous progress has been made over the past quarter-century studying the genetics of gametogenesis and the resulting gametes and embryos. Studies merging molecular techniques and conventional cytogenetics are now beginning to bridge the gap between what we have learned about the meiotic process in males and females and what we know of the mitotic chromosomes of zygotes. Numerical abnormalities in sperm, oocytes and embryo can now diagnosed by fluorescence in situ hybridization (FISH). "At risk" couples can, therefore, have only unaffected embryos replaced in the sterus and avoid the possibility of terminating a pregnancy that might only be diagnosed as affected later gestation. Single-cell genetic analysis has also provided powerful tools for studying genetic defects arising during early human development. Recent studies of sperms, oocytes and cleavage-stage human embryos have revealed an unexpectedly high incidence. These genetic abnormalities are likely to contribute to early pregnancy loss and have important implications for improving pregnancy rates in infertile couples by assisted reproduction. The widespread use of preimplantation genetic diagnosis (PGD) awaits further documentatio of safety and accuracy. Other issues also must be addressed. First, the ethical issues regarding germ cell and embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for non-genetic disorders such as gender selection. Finally, the experimental nature of these procedure must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved assisted reproductive technology success rates may make PGD a more widely use clinical tool. The future awaits these development.velopment.

• Development and Reproduction
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• v.20 no.4
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• pp.305-314
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• 2016

The marine medaka, Oryzias dancena is a suitable sample as a laboratory animal because it has a small size and clearly distinguishes between female and male. Data on the growth and maturity of the diploid and triploid sea cucurbit species suitable for laboratory animals are very useful for studying other species. Triploidy was induced in the marine medaka by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 45 min, applied two minutes after fertilization. The diploid and triploid male fish were larger than their female counterparts (P<0.05), and the concentrations of thyroid stimulating hormone (TSH) and thyroxine (T4) were higher in the induced triploids over 1 year (P<0.05). In both the diploid and tri-ploid groups the concentrations of TSH and T4 were higher in the male fish than in the females (P<0.05), while the testo-sterone and estradiol-$17{\beta}$ concentrations in the induced triploids were lower than in the diploids (P<0.05). The gonadosomatic index (GSI) of the triploid fish was lower than that for the diploids, and the GSI for females in each ploidy group were higher than that for the males. For both groups the GSI was highest at 4 months of age, and decreased thereafter to 12 months. Analysis of the gonads of one-year-old triploid fish suggested that the induction of triploidy probably causes sterility in this species; this effect was more apparent in females than in males.

• Development and Reproduction
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• v.23 no.1
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• pp.35-42
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• 2019

Fish shows great difference in growth rate between individuals during larval development and early growth. This difference seriously reduces the production efficiency in fish culture. Growth hormone (GH)/Insulin-like growth factor 1 (IGF1) system is said to play some pivotal roles in fish growth. In this study, we investigated differences of GH, IGF1 and GHR gene expressions in juvenile red spotted grouper (Epinephelus akaara) with different growth performance. Red spotted groupers were reared under the same environmental condition (water temperature $24{\pm}1^{\circ}C$, natural light) for 96 days after hatching. They were divided into 3 groups by size (fast growing, middle growing and slow growing groups: FGG, MGG, and SGG, respectively). RNA was extracted from the brain, liver and muscle tissues from each group, and target gene expression was examined by real-time PCR. In the brain with pituitary gland, expression of GH gene in FGG was significantly higher than the expression in SGG, but the expression of IGF1 and GHR genes in the muscle was highest in SGG. Difference of GHR and IGF1 mRNA in the liver between groups with different growth performance was less clear than that in other tissues, although level of IGF1 mRNA was higher in SGG than in MGG. These results suggest that hormonal governing of growth is not the same in fast growing and slow growing fish, and size grading could cause a shift of hormonal state and growth pattern in this species.