• Development and Reproduction
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• v.21 no.1
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• pp.19-33
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• 2017

Optimum concentrations of anesthetic clove oil and anesthetic lidocaine-HCl were determined for a species of adult marine medaka, Oryzias dancena, over a range of salinity conditions, and investigated in a transport simulation experiment by analyzing various water and physiological parameters. Research indicated that the higher the concentration of anesthetic at each salinity, the shorter the anesthesia time at each salinity. At each concentration, fish were anesthetized slower at water salinities over 10 ppt (P<0.05). Anesthesia time at 10 ppt was faster than any other salinity. In 10 ppt salinity, the dissolved oxygen (DO) concentrations and respiratory frequencies of the clove-oil-administered groups decreased until 48 hours (P<0.05), whereas the $NH_4{^+}$ and $CO_2$ concentrations increased until 48 hours (P<0.05). In same period, the DO, $NH_4{^+}$, and $CO_2$ concentrations and respiratory frequencies all decreased as the clove oil concentration increased (P<0.05). The trends in the DO, $NH_4{^+}$, and $CO_2$ concentrations and respiratory frequencies in the lidocaine-HCl-administered groups were similar to those in the clove-oil-administered groups. In conclusion, clove oil and lidocaine-HCl are effective anesthetics, improving the transportation of the marine medaka. The results from this study will contribute to safe laboratory handling of the marine medaka, which are commonly required by many research studies and experiments.

• Development and Reproduction
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• v.21 no.1
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• pp.101-110
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• 2017

On June 14, 2008 (the first experiment) and July 24, 2008 (the second experiment), the shores of the Boseong River and the sandy beaches, Seokgok-myun, Moksadong-myun, Gokseong-gun in Jeollanam Province were investigated and a total of 29 soft-shelled turtle (Tryonyx sinensis) eggs in the natural spawning nest eggs were collected (13 eggs were collected in the first experiment and 16 eggs in the second experiment). The temperatures in the natural spawning nests were $25.9-36.9{\pm}0.5^{\circ}C$, the depth of the eggs was $5.2-7.5{\pm}0.5cm$ as the distance of the average $6.4{\pm}0.5cm$. 29 eggs were scattered at least 0.2 cm interval. Artificial incubation of 29 eggs was conducted in artificial nest boxes in thermo-plastic composition of the incubator, and then incubated at $26.5-35.5{\pm}0.5^{\circ}C$, and an average constant temperature was $31.2-32.1{\pm}1.0^{\circ}C$. The incubation days ranged from 53 to 55. In case of most turtles, incubation at $31^{\circ}C$ (higher temperatures) generally produces all or mostly females, while incubation at $25^{\circ}C$(cooler temperatures) produces all or mostly males. Exceptionally, in case of genus Trionyx, the sex ratio of female : male of T. sinensis of a freshwater soft-shelled turtle was approximately 1:1, which differs from other genera of turtles and makes T. sinensis Strauch only turtles presently known to lack temperature-dependent sex determination.

• Development and Reproduction
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• v.21 no.2
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• pp.181-191
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• 2017

A comparison of the growth, hematological values, fatty acids, and gonadal and growth hormonal changes of river puffer, Takifugu obscurus, tiger puffer, T. rubripes, their hybrids (river puffer ${\times}$ tiger puffer) and hybrid triploids was performed during 3 months of their early growth period. Several features were observed during these 3 months: hybrids showed the highest levels of specific growth rate, 1.48%; hybrid triploids showed the smallest change in viscera fat (P<0.05), but GSI was not significantly different among groups (P>0.05). Considering hematological parameters, hybrid triploids had increased mean corpuscular volume and mean corpuscular hemoglobin (P<0.05), but other parameters were not significantly different between groups (P>0.05). With respect to fatty acids, puffer fish, hybrids and hybrid triploids contained fatty acids such as SFAs, MUFAs, n-3 PUFAs and n-6 PUFAs. There were significantly different amounts of total fatty acids between groups (P<0.05), however, rates of changes in fatty acids did not differ significantly between groups (P>0.05). Gonadal hormone (estradiol and testosterone) changes in the river puffer and tiger puffer were significantly higher than that observed in hybrids and hybrid triploids. The hybrids and tiger puffers had higher amounts of growth hormone (thyroid stimulating hormone and thyroxine) than the hybrid triploids and river puffers (P<0.05).

• Fisheries and Aquatic Sciences
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• v.19 no.9
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• pp.41.1-41.7
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• 2016

Background: The transgenic approach using estrogen-responsive regulator in fish has been given much attention as a potential means to detect and/or address estrogen-related aquatic pollutions. In order to address the development stage- and reproduction status-dependent expression patterns of the chgH-rfp transgene (red fluorescent protein transgene driven by choriogenin H promoter) in marine medaka Oryzias dancena, naturally occurring red fluorescent protein (RFP) signals under non-exposed conditions as well as the transgenically induced RFP signals under estrogen-exposed conditions were assayed. Results: Female transgenics begun to show naturally occurring RFP signals from the age of 7 weeks post hatching (WPH) without experimental estrogen exposure. Afterward, these RFP signals in female transgenics became robust with the progress of ovarian maturation. On the other hand, male transgenics did not show any naturally occurring RFP signal under non-exposed conditions irrespective of developmental stages and maturation statue. Upon exposures using estradiol-$17{\beta}$ (E2) and $17{\alpha}$-ethinylestradiol (EE2), RFP signals were significantly induced specifically in the livers of transgenic males. Conclusions: Male chgH-rfp transgenics were able to keep the "off" state of RFP expression during their entire life cycle unless exposed to exogenous estrogens. Owing to their tight regulation capability of estrogen-responsive transgene, transgenesis of chgH-rfp in male marine medaka could offer a useful model system for future ecotoxicogenomic studies regarding estrogenicity-related issues in aquatic and marine environments.

• Development and Reproduction
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• v.18 no.1
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• pp.33-41
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• 2014

We investigated the process of yolk absorption in Korean rose bitterling, Rhodeus uyekii, and determined the changes in its morphometric characteristics. The R. uyekii from 1 days post hatching (DPH) to 21 DPH, the eye diameter (ED) was 5.4 at 5 DPH. Thereafter, the ED/total length (TL) ratio increased to 10.7 at 21 DPH (p<0.05). The yolk length (YL) decreased from 95.4 to 1.1 by 21 DPH, and this rate of decrease was greater than that for any other dimension (p<0.05). 12 morphometric dimensions/TL for the R. uyekii were measured at each sampling day from 21 DPH to 170 DPH. At just hatching, the average TL and BW were $6.1{\pm}0.09mm$ and $4.9{\pm}0.07mg$, respectively. At 53 DPH, the average TL was $12.9{\pm}0.28mm$ and the average BW was $14.7{\pm}0.72mg$; the total length growth equation was $TL=5.507e^{0.038t}$ ($R^2=0.916$). Further, the body weight growth equation was $BW=3.3647e^{0.0296t}$ ($R^2=0.9354$). The dimensions with regard to body depth showed the greatest growth rates in the external characteristics of the fish (p<0.05). The patterns of the morphometric characteristics measured in this study can be classified in three ways. The patterns were shown to be increased (y=0.0992x+12.07, $R^2=0.8333$), decreased (y=-0.0569x+42.029, $R^2=0.8395$) or maintained (y=0.005x+18.263, $R^2=0.3678$) from 21 DPH to 170. These results will provide useful indices for the successful rearing of the R. uyekii.

• Development and Reproduction
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• v.18 no.1
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• pp.13-23
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• 2014

This study is conducted to monitor the morphological developmental features of the egg development, larvae and juvenile of Epinephelus septemfasciatus, the fertilized eggs were gotton using artificial insemination. Matured parents are collected from marine caged fish farms in Geomun-ri, Samsan-myeon, Yeosu-si, Jeollanamdo Korea in June 2012. The fertilized eggs were pelagic eggs containing one oil globule, and measured 0.81~0.89 mm ($0.85{\pm}0.04mm$, n=50) in diameter. In regard to rearing environment, the water temperature is $21.0{\sim}23.0^{\circ}C$ and the salinity is 32.0~33.2‰. Hatching was observed from 48 hours after fertilization, the mouth and anus of prelarvae was not opened but had egg yolk at newly hatched. 4 days after hatching, the mouth and anus of postlarvae was opened and began to eat Rotifer and was measured 2.40~2.49 mm ($2.45{\pm}0.03mm$ n=10) in total length. 12 days after hatching, postlarvae was measured 3.77~4.67 mm ($4.27{\pm}0.33mm$) in total length, its the second pole tide of dorsal fin and the first pole tide of pelvic fin was extended longitudinally. 71 days after hatching, juvenile was measured 40.5~45.4 mm ($42.6{\pm}2.04mm$) in total length. Seven bands were observed in body, and pole tides of dorsal and pelvic fins were shortened.

• Development and Reproduction
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• v.17 no.1
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• pp.55-62
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• 2013

Puberty is the developmental period which animals obtain the ability of reproducing sexually for the first time in life. In commercially important aquaculture fish species, the onset of puberty is a matter of major interest due to controlling of sexual maturation to improve broodstock management. To investigate pubertal characteristics of female longtooth grouper (Epinephelus bruneus), specimens were classified into three groups by the bodyweight, including 1, 2, and 3 kg group. Thereafter, we focused on ovarian development and level changes of endocrine regulation factors (GnRH, GTHs, steroid hormone). In the non-breeding season (April), the levels of endocrine regulation factors showed increasing trends in accordance with bodyweight gaining; nevertheless, the oocytes were growth phase belongs to almost peri-nucleous stages in all groups. In the breeding season (June), the levels of endocrine regulation factors were fluctuated that decreases in levels of sbGnRH and $FSH{\beta}$ mRNA expressions along with serum $E_2$ concentrations in 3 kg of group. However, $LH{\beta}$ mRNA expression levels sustained increasing trends by the bodyweight. Moreover, the oocytes developed that 2 kg and 3 kg groups obtained plentiful vitellogenic oocytes while 1 kg group was still composed with greater part of pre-vitellogenic oocytes. Especially, the oocytes of 3 kg group reached over 450 ${\mu}m$ of diameters that indicating possibility to enter the final maturations. These results suggest that the progress of pubertal development in female E. bruneus could be classify into three phases via bodyweight, including pre-puberty (1 kg), early-puberty (2 kg) and puberty (3 kg).

• Development and Reproduction
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• v.23 no.4
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• pp.323-331
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• 2019

In this study, an experiment was conducted to investigate the annual reproductive cycle of a Korean flat bittering, Acheilognathus rhombeus, from Ogok-myeon located in Seomjin River. The reproductive cycle was examined histologically regarding water temperature and day length of the habitat, the gonadosomatic index (GSI), and developmental characteristics of female and male gonads. The maximum GSI was found to be 3.50±0.53 and 1.36±0.14 for females and males, respectively, when the water temperature and day light was 16.9℃ and 11.3 hours, respectively in October 2018. On the other hand, the minimum GSI was found to be 0.16±0.09 and 0.69±0.15 for males and females in December 2018 and February 2019, respectively. The ovipositor of females appeared from August to November 2018. We compared and calculated the stages of germ cell developmental characteristics in the testis and ovaries to determine the reproductive cycle. According to the result, we classified the female A. rhombeus reproductive cycle into four phases, which are ripe and spawning phase (October), degenerative phase (November to December), growing phase (January to March) and mature phase (April to September). The annual reproductive cycle of male A. rhombeus was categorized into four phases: mature phase (June to October), degenerative phase (November to March), resting phase (April) and growing phase (May). The Korean flat bittering is an autumn-spawner as the main spawning season in October. In male, testicular spermatogonia appeared all year-round, and the ripe and releasing phase, which is characteristics of the spawning season in other fish, did not appear.

• Development and Reproduction
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• v.23 no.4
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• pp.367-375
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• 2019

Pufferfish (Takifugu spp.) are economically important edible marine fish. Mistakes in pufferfish classification can lead to poisoning; therefore, accurate species identification is critical. In this study, we used the mtDNA cytochrome c oxidase subunit I gene (COI) to design specific primers for six Takifugu species among the 21 domestic or imported pufferfish species legally sold for consumption in Korea. We rapidly and simultaneously identified these pufferfish species using a highly efficient, multiplex polymerase chain reaction (PCR) system with the six species-specific primers. The results showed that species-specific multiplex PCR (multiplex species-specific polymerase chain reaction; MSS-PCR) either specifically amplified PCR products of a unique size or failed. MSS-PCR yielded amplification fragment lengths of 897 bp for Takifugu pardalis, 822 bp for T. porphyreus, 667 bp for T. niphobles, 454 bp for T. poecilonotus, 366 bp for T. rubripes, and 230 bp for T. xanthpterus using the species-specific primers and a control primer (ca. 1,200 bp). We visualized the results using agarose gel electrophoresis to obtain accurate contrasts of the six Takifugu species. MSS-PCR analysis is easily performed and provides identification results within 6 h. This technique is a powerful tool for the discrimination of Takifugu species and will help prevent falsified labeling, protect consumer rights, and reduce the risk of pufferfish poisoning..

• Development and Reproduction
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• v.19 no.4
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• pp.209-215
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• 2015

Red spotted grouper, Epinephelus akaara is a popular aquaculture species in many Asian countries. This species is a protogynous hermaphrodite that first differentiates into female and changes to male later. Due to this reproductive characteristic, stable supply of male and female gametes is a key to the success of seed production in this species. Thus, understanding early gonadal differentiation is required to develop effective sex control techniques. Red spotted grouper were reared in indoor tanks and sampled every 5 days from 40 days post-hatch (DPH) to 130 DPH. Changes of gonadal tissues were examined and analyzed by means of histology. A pair of gonadal primordium has already existed underneath the kidney in the posterior part of the body cavity at 38 DPH when this study began. Gonadal primordia of 38, 40 DPH consisted of germ cells surrounded by a few somatic cells. The blood vessel was observed in the gonadal primordium at 45 DPH. The number of somatic cells and size of gonadal primordium increased age-dependently up to 60 DPH. Formation of ovarian cavity was obvious by two protuberant aggregations of somatic cells at 65 DPH. Completed ovarian cavity and oogonia were first observed in the gonad of one fish sample at 105 DPH. Based on these histological observations, it can be suggested that induction of primary male differentiation could be more successfully applied at around 60 DPH in this species.