• Korean Journal of Environmental Biology
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• v.36 no.3
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• pp.352-358
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• 2018

This study aimed to develop a species identification method for the egg and fry of the three Korean bitterling fishes (Pisces: Acheilognathinae), including Acheilognathus signifer, Acheilognathus yamatsutae and Rhodeus uyekii based on the PCR-based Restriction Fragment Length Polymorphism (RFLP) markers. We conducted a field survey on the Deokchicheon River from the North Han River basin, where the three Acheilognathinae species co-occur, and also analyzed the existing sequence dataset available from the GenBank. We found coexistence of the three species at the study site. The egg and fry were obtained from the host mussels (Unio douglasiae sinuolatus) by hand from May to June 2015 and in May 2017. To develop PCR-based RFLP markers for species identification of the three Acheilognathinae fish species, restriction enzymes pinpointing species-specific single nucleotide variation (SNV) sites in mitochondrial DNA COI (cytochrome oxidase I) and cyt b (cytochrome b) genes were determined. Genomic DNA was extracted from the egg and fry and RFLP experiments were carried out using restriction enzymes Apal I, Stu I and EcoR V for A. signifer, A. yamatsutae and R. uyekii, respectively. Consequently, unambiguous discrimination of the three species was possible, as could be seen in DNA band patterns from gel electrophoresis. Our developed PCR-based RFLP markers will be useful for the determination of the three species for the young and would assist in studying the spawning patterns and reproductive ecology of Acheilognathinae fishes. Furthermore, we believe the obtained information will be of importance for future maintenance, management and conservation of these natural and endangered species.

• Korean Journal of Environment and Ecology
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• v.33 no.2
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• pp.202-215
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• 2019

This study was conducted to investigate the spawning preference of the Acheilognathinae fishes in relation to the shell size of host mussels after identifying the species of eggs and fries in the host mussel using our recently developed RFLP (Restriction Fragment Length Polymorphism) molecular marker at four sites [Hongcheon Naechoncheon (HN) and Deokchicheon (HD) from the North Han River basin and Jeongseon Goljicheon (JG) and Joyanggang (JJ) from the South Han River] in South Korea during May in each year between 2015 and 2018. The Acheilognathinae fish observed in the studied sites included one species (Acheilognathus signifer) in HN and JG, three species (Rhodeus uyekii, A. signifer, and Acheilognathus yamatsutae) in HD, and two species (A. signifer and Acheilognathus yamatsutae) in JJ, and we collected 982 host mussels (Unio douglasiae sinuolatus) that inhabited in all four sites. Using the RFLP molecular marker, we confirmed 46 eggs and fry of the Acheilognathinae fish (454 A. signifer, 43 Acheilognathus yamatsutae, and 149 Acheilognathus yamatsutae) in Unio douglasiae sinuolatus (N=163; 16.6%). We compare the average shell length, shell height, and shell width of mussels with [presence] eggs/fry and mussels without [absence] eggs/fry to examine the spawning preference according to the size of host mussels in each site. The results show that the shell length (1.98 mm), shell height (0.85 mm), and shell width (0.73 mm) of mussels with the eggs/fry were significantly larger (Mann-Whitney U test, P=0.002; difference=1.98 mm) than those of mussel without eggs/fry in HD where three species cohabitated. Although the shell length, shell height, and shell width of mussels with the eggs/fry were larger also in the other three sites, the difference was not statistically significant. In addition, we analyzed the mean number of spawned eggs and fry of each species and found $9.31{\pm}5.94$ R. uyekii, $2.86{\pm}2.45$ A.signifer, and $2.50{\pm}1.32$ A. yamatsutae. R. uyekii spawned 6.45-6.81 more eggs than A.signifer and A. yamatsutae on average per mussel, and it was statistically significant (Kruskal-Wallis test, P < 0.001). These findings indicate that the three species of Acheilognathinae fish tend to prefer larger mussels as their spawning hosts, and this tendency increases when the number of cohabitating bitterling fish species increases. Moreover, A.signifer and A. yamatsutae spawned a smaller number of eggs evenly in more host mussels while R. uyekii spawned many eggs on relatively fewer mussels. We found mussels (N=4) having the eggs/fry of two coexisting species, A. signifier and A. yamatsutae in HD and JJ where more than two bitterling fish species occurred. It suggests the interspecific competition taking place between the Acheilognathinae fishes for utilizing the same resource of mussels for spawning when two or more species cohabitate. This study is expected help to understand better the spawning patterns and reproductive ecology of the Acheilognathinae fishes, which will provide insightful information for advancing our understanding of their ecological relationships - mutualism or host-parasitism - with host mussels.

• Proceedings of the Korean Aquaculture Society Conference
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• 2006.05a
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• pp.566-567
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• 2006

• The Science & Technology
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• no.11 s.438
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• pp.50-53
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• 2005

몸이 세로로 납작한 소형 민물고기인 납자루 아과(Acheilognathinae) 어류는 잉어과에 해당하며 전세계에 40여 종이 알려져 있다. 우리 나라에는 이미 멸종된 서호납줄개를 포함하여 납줄개속, 납자루속 및 큰납지리속의 3속 14종이 있으며, 그 중 8종은 우리 나라의 고유종이다. 수컷은 화려한 혼인색을 내고 암컷은 산란관을 내어 민물조개에 산란하는 특성이 있다.

• Korean Journal of Ichthyology
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• v.25 no.2
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• pp.74-81
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• 2013

Factors influencing spawning or respawning conditions of the bitterling, Acheilognathus signifer were investigated in the aquariums. Inducing the female spawning was more sensitive to the mussel than the male. The ovipositor of the female was periodically elongated and reduced from 4 to 6 days with the presence of mussel. During of the elongated state of the ovipositor was 1 to 2 days. As the result of natural spawning in the aquariums, it takes 1 to 3 days (mean 1.6) to spawn. The most important factor to respawn spawned females again was the mussel. It takes 11 to 53 days (mean 29.5) from extraction to reextraction of females which have elongated ovipositor. The egg numbers of reextraction were 2 to 41 (mean 19). This experiments showed the tendency the more standard length of females grows, the more the number of extracted eggs increases.

• Korean Journal of Ichthyology
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• v.18 no.3
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• pp.266-272
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• 2006

The egg development and early life history of Korean endemic bitterling Rhodeus pseudosericeus were observed under the controlled water temperature, $17.0{\pm}0.5^{\circ}C$. Fertilized eggs are opaque yellow in color, measuring 2.9~3.1 mm in length and 1.6~1.9 mm in breadth. The eggs of this species began to hatch about seventy hours after insemination and the range of total length of larvae were 3.2~3.4 mm. The larvae reached at the heterotrophic stage about twenty-five days after hatching. R. pseudosericeus had a pair of wing-shaped yolk projection during the prelarval stage that is a common character of the genus Rhodeus.

• Korean Journal of Ichthyology
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• v.27 no.2
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• pp.78-85
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• 2015

Population ecology of Rhodeus uyekii (Pisces: Acheilognathinae) was investigated at the Bongseocheon, Mankyeonggang (River) in Korea, from January to December 2014. It mainly inhabits the slow-flowing waters having mud bottoms. Measurement of the standard length of the population indicated that the following standardization: 24~29 mm are one year old, 30~35 mm are two years old, 36~39 mm are three years old, and >40 mm group are regarded over four years old (October in 2014). The spawning season was from March to June, with water temperatures in the range $11.5{\sim}27.0^{\circ}C$. The sex ratio of female : male was 1 : 0.77. The number of eggs in the ovaries was 104~168 ($mean{\pm}SD$: $124{\pm}18.1$). The size of the matured eggs was 2.8~3.5 ($3.2{\pm}0.15$) mm in the long axis, and 1.4~1.5 ($1.4{\pm}0.03$) mm in the short axis. The eggs were found in Unio douglasiae and the number of eggs, larvae or both was 3~18 ($10.2{\pm}5.0$). Stomach contents were mainly phytoplanktons such as the genera Navicular, Cocconeis, Cymbella (Bacilariophyceae) and Cloniophora (Chlorophyceae).

• Korean Journal of Ichthyology
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• v.29 no.4
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• pp.235-243
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• 2017

Reproductive characteristics of Rhodeus pseudosericeus (Pisces: Acheilognathinae) was investigated at the Heukcheon (River), Yangpyeong-gun, Gyeonggi-do, Korea from January to July in 2012. This fish prefers heavily vegetated stagnant pool with Phragmites japonica and Salix koreensis and area with high proportions of sand and mud. The standard length of the population indicated that 13~22 mm group is 0-year old, 30~45 mm group is 1-year old, 45~55 mm group is 2-year old and 45~70 mm group is more than 3-year old (June in 2012). The sex ratio of female to male was 1 : 0.80. The spawning season was from March to June with water temperatures in the range $11.0{\sim}26.2^{\circ}C$. The embryos were found with no egg mass in Unio douglasiae sinuolatus and the number of those was 1~22 ($5.8{\pm}5.51$). The number of the embryos found in each of the four parts of the mussel gill was 1~9 ($2.7{\pm}2.16$) in left outer, 1~2 ($1.5{\pm}0.71$) in left outer, 1~5 ($1.8{\pm}1.79$) in left outer and 1~22 ($5.8{\pm}5.51$) in left outer.

• Korean Journal of Environment and Ecology
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• v.35 no.2
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• pp.115-125
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• 2021

This study surveyed the population ecology of Acheilognathus chankaensis (Pisces: Acheilognathinae) in the vicinity of the Churyeongcheon (Stream) and Seomjingang (River) in Jeonbuk, Korea, from February 2006 to May 2007. The survey found the habits mainly in the slow-flowing waters having sand, mud, and gravel bottoms. Age groups estimated by the standard length-frequency distribution of the population indicated that the one-year-olds were 30-46 mm long, the two-year-olds were 47-58 mm long, the three-year-olds were 59-68 mm long, and the four-year-olds were longer than 69 mm. The gonadosomatic index (GSI) and the ratio of ovipositor length/standard length were the highest in March (♀: 12.9%, ♂: 5.0%) and in April (46.3%), respectively. The ratio of mature egg diameters that were 1.6 mm was highest in May (13.2%). The comprehensive analysis of spawning characteristics showed that the spawning season was from March to June with water temperatures in 11.5-22.5℃, and the sex ratio of female:male was 1:1.13. The fecundity in the ovaries and the number of mature eggs were 318±174.5 (50~583) and 220±139.2 (50~406), respectively. The size of the mature eggs was 2.04±0.110 mm on the long axis and 1.55±0.100 mm on the short axis for the study period. Gut contents of A. chankaensis were mainly phytoplankton such as the genera Fragilaria, Synedra, Navicular, Cymbella (Bacillariophyceae), and Cosmarium (Chlorophyceae).

• Korean Journal of Ichthyology
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• v.17 no.2
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• pp.105-111
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• 2005

Most of the eggs and (or) pre-larvae of Acheilognathus signifer were observed from the gills of Unio douglasiae sinuolatus, 30~45 mm in the shell length, that is the host mussel of A. signifer. There was no selectivity in proportion to mussel size at the range observed. One to seven individual eggs and (or) pre-larvae were found in the mussels, with a mean of 2.5 individuals, and the rate of possessing one egg and (or) pre-larva was 50.0%. When the prelarva of A. signifer acquired swimming ability inside the mussel, it moved into the suprabranchial chamber. It was estimated that the growth period was 4~6 weeks. The minute tubercles of the pre-larvae were observed immediately after hatching. Absorption of the minute tubercles was observed starting the 7 th day, with most of tubercles absorbed 13 days after hatching and completed 20 days after the yolk was entirely absorbed.