• 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.

• Korean Journal of Ichthyology
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• v.26 no.4
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• pp.288-294
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• 2014

Host mussel utilization for spawning of the oily shinner, Sarcocheilichthys variegatus wakiyae (Pisces: Cyprinidae), from Korea was investigated in part of the Dalcheon (river) in Cheongcheon-myeon, Goesan-gun, Chungcheongbuk-do, Korea from April to June 2011, 2013. The non-synchronicity of mature and immature eggs obtained from females of S. v. wakiyae and the number of spawned eggs found was less than those in the mussels, suggesting that these fish may be batch spawners. Five species of freshwater mussels - Unio douglasiae sinuolatus, Lanceolaria grayana, Anodonata arcaeformis flavotincta, Lamprotula leai, Corbicula fluminea - were found at the survey area (wild conditions) but three species of freshwater mussels - A. a. flavotincta (oviposition rate, 50.0%), U. d. sinuolatus (16.2%), L. leai (11.1%) - were host of S. v. wakiyae. Spawning of S. v. wakiyae occurred mainly in the mantle cavity (wild conditions, 77.4%; experiment, 62.8%) of the mussels that were connected more to the inhalant siphon than the suprabranchial cavity (22.6%; 31.8%) that linked to the exhalant siphon. Bitterlings prefer to lay eggs in L. leai and S. v. wakiyae prefers A. a. flavotincta as a spawning host. These were considered to be the result of niche partitioning.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.6
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• pp.581-585
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• 1985

This study was carried out with blue mussels at Otsuchi Bay in Japan from August 1981 to May 1982, in order to investigate the effect of the parasitic pea crab (Pinnotheres pholadis) on the reproductive capacity of host blue mussels (Mytilus edulis galloprovincialis). The results show that pea crabs delay gonad development of host mussels about one month and lower their fatness rate by intercepting nutrients. The parasitization with pea crabs is thought not to prevent spawning activity itself, but to reduce the number of germ cell resulting in the reduced space of gonads.

• Korean Journal of Ichthyology
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• v.25 no.4
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• pp.208-215
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• 2013

Host species preference of Acheilognathus signifer for spawning in freshwater mussels was investigated at the part of the Dalcheon Stream (site 1) and the Gaedaecheon Stream (site 2) in Chungcheongbuk-do, Korea from April to June (spawning period) 2011. The spawned mussel rate of A. signifer at the study site 1 (wild condition) was 63.6% in Lamprotula leai and 2.1% in Unio douglasiae sinuolatus. It was not found in Anodnta arcaeformis flavotincta and Lanceolaria grayana. The number of eggs, larvae, or both of A. signifer at the study site 1 (wild condition) was 1~41 ($12.0{\pm}9.21$) in L. leai and 6 in U. d. sinuolatus. As a result of experiment 1, A. signifer was only spawned in L. leai with 60% of spawned mussel rate and the number of eggs, larvae, or both was 1~19 ($6.8{\pm}5.44$). According to experiment 2, A. signifer was spawned in all mussels. Spawned mussel rate of A. signifer was 75.0% in L. leai, 42.9% in U. d. sinuolatus and 21.4% in A. a. flavotincta and the number of eggs, larvae, or both was 1~35 ($13.0{\pm}8.80$) in L. leai, 1~26 ($5.2{\pm}6.92$) in U. d. sinuolatus and 2~19 ($8.6{\pm}5.64$) in A. a. flavotincta. Experiment 3 showed A. signifer was solely spawned in L. leai and the number of egg was from 2 to 10 ($6.7{\pm}4.16$). This study confirmed A. singifer is not so much generalist as specialist. We found the most preference species for spawning in the mussels by A. signifer was L. leai.

• The Korean Journal of Ecology
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• v.28 no.4
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• pp.193-198
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• 2005

This study investigated the characteristics of the eggs and yolk sac larvae of Korean striped bitterling, Acheilognathus yamatsutae, spawned and grown In mussels. The number of eggs in the ovary was small ($358{\pm}108$ SD). The eggs were oval and large, and the formation of the perivitelline space was narrow. The eggs were hatched at only 41 hours after fertilization but the hatched larvae were underdeveloped. The development of yolk projection and minute tubercles on the skin surface was notable, along with the vividly moving tail in the hatched larvae. The yolk projection and minute tubercles were disappeared upon enhancement of the motor ability of the larvae was enhanced. The formation of eyes and body pigments of the larvae was relatively delayed in comparison with that of other cyprinid larvae. After completely consuming the yolks the larvae escaped from the mussel for free swimming and exogenous feeding.

• Korean Journal of Ichthyology
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• v.32 no.2
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• pp.63-69
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• 2020

This study investigated spawning characteristics of Hangang bitterling, Rhodeus pseudosericeus inside the host mussel, Unio douglasiae sinuolatus at the Heukcheon Stream in Korea from April to June 2017. 327 mussels were collected, among them, 34 (14.1%) were spawned mussels of R. pseudosericeus with glochidia. The mean number of R. pseudosericeus embryos with or without glochidia by the mussel gill position was 5.0±3.58 (range, 1~8) vs. 2.8±2.13 (1~9) in the left outer demibranch, 1.5±0.96 (1~3) vs. 1.7±0.82 (1~3) in the left inner demibranch, 1.4±0.51 (1~2) vs. 2.1±1.85 (1~6) in the right inner demibranch, and 6.4±4.82 (1~20) vs. 3.0±1.86 (1~6) in the right outer demibranch. The frequency of R. pseudosericeus embryos with or without glochidia by the mussel gill position was 82.6% vs. 73.4% in the left outer demibranch, 17.4% vs. 9.4% in the left inner demibranch, 26.1% vs. 15.6% in the right inner demibranch, and 58.7% vs. 67.2% in the right outer demibranch. The mean number of R. pseudosericeus embryos with or without glochidia was 8.5±6.43 (1~23) vs. 4.5±3.38 (1~14). The number and frequency of R. pseudosericeus embryos were higher in the mussel with glochidia and the outer demibranchs than those without glochidia and with inner demibranchs.

• Korean Journal of Environmental Biology
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• v.41 no.3
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• pp.229-246
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• 2023

The bitterling (Cyprinidae, Acheilongnathinae) is a temperate freshwater fish with a unique spawning symbiosis with host mussels. Female bitterlings use their extended ovipositors to lay eggs on the gills of mussels through the mussel's exhalant siphon. In the present study, in April of 2020, we investigated spawning frequencies and patterns of three bitterling fish species in host mussel species in the Nakdong River basin (Hoecheon). During field surveys, a total of four bitterling and three mussel species were found. We observed bitterling's spawning eggs/larvae in the three mussel species: Anodonta arcaeformis(proportion spawned: 45.5%), Corbicula fluminea(12.1%), and Nodularia douglasiae (45.2%). The number of bitterlings' eggs/larvae per mussel ranged from 1 to 58. Using our developed genetic markers, we identified the eggs/larvae of each bitterling species in each mussel species (except for A. macropterus): A. arcaeformis (spawned by Acheilognathus yamatsutae), C. fluminea (A. yamatsutae and Tanakia latimarginata), and N. douglasiae (A. yamatsutae, Rhodeus uyekii, and T. latimarginata). Approximately 57.6% of N. douglasiae mussel individuals had eggs/larvae of more than one bitterling species, suggesting that interspecific competition for occupying spawning grounds is intense. This is the first report on bitterling's spawning events in the Asian clam C. fluminea from Korea; however, it should be ascertained whether bitterling's embryo undergoes successful development inside the small mussel and leaves as a free-swimming juvenile. In addition, the importance of its conservation as a new host mussel species for bitterling fishes needs to be studied further.

• 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 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.

• Korean Journal of Ichthyology
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• v.6 no.1
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• pp.51-59
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• 1994

Morphology of glochidia of two species of Unionidae mussels, Unio douglasiae and Anodonta woodiana were studied based on SEM, and their attachment on Acheilognathus yamatsutae(Acheilognathinae) was observed in the experimentally infected fishes. The glochidia were round-triangle in shape and possessed larval threads, hooks, hooklets, and sensory hairs. Glochidal attachment and cyst formation of the fish were observed without regard to feeding habit and habitat. Density of glochida on the fish (Acheilognathus yamatsutae) was in the order of pectoral fin, dorsal fin, gill and caudal fin. The host fish became encysted about 5-10 hours to the glochidia, and the glochidia detached about 5-6 days after attachment at water temperature $26{\pm}1^{\circ}C$.