• 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.18 no.1
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• pp.27-35
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• 2006

During spawning season of female in a Korean oily bitterling, Acheilognathus koreensis, the ventral region near the base of the pectoral fin becomes to be protruded outward of body and enlarged. This ventral process consists of both organs as rectum (vent) and inner ovipositor. The rectum consists of mucosa, lamina propria-submucosa, muscularis, and squamous epithelial layer (peritoneum=serosa) surrounding them. The mucosa contains numerous mucous cells meaning acid mucopolysaccharides in nature. The inner ovipositor is similar to that of the rectum, but the mucosa have no mucous cell, unlike that of the rectum. Whereas, the outer ovipositor has a straight and long tube which are not connected with the ventral process any more. The outer ovipositor was similar to the structure of the inner ovipositor in the ventral process. However, the outer ovipositor has no muscularis, and consists of three layers: mucosa, lamina propria-submucoa, and squamous epithelia. The outer ovipositor without the muscularis seems serves as a tube that eggs discharged from the outer ovipositor allow to send inside mussel, unlike that of the inner ovipositor performing rhythmic contractions of the layers of the muscularies for propelling to the matured oocytes to the outer ovipositor.

• 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 Ichthyology
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• v.2 no.2
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• pp.169-181
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• 1990

The development of eggs and larvae, and minute scale-like tubercles on the skin surface of larval Acantlaorhodeus gracilis from Korea were observed. They spawned from late March through the middle of June. A fish spawned at least 5 times and the number of eggs averaged 304 per oviposition. Unfertilized eggs are nearly ovoid-shaped(mean$\pm$SD=$2.09{\pm}0.04\;mm$ in length with range of 2.05 to 2.13 mm and mean$\pm$SD=$1.26{\pm}0.02\;mm$ in breadth with range of 1.24 to 1.30 mm measured for the 50 eggs) and opaque yellow color. The tip of egg membrane at the animal pole side swelled and formed a few hilly projections. The shape of the eggs was just like a loquat. Most of embryos began to hatch out in thirty-eight hours after insemination at $22{\pm}1^{\circ}C$ in water temperature. As regards the morphological characters of the eggs and larval development, Acan. gracilis was similar to Acan. asmussi, Acheilognathus rhombeus, A. longipinnis and Pseudoperilampus typus. The larvae of this species is unique particularly in the following two characters, i.e., 1) scale-like tubercles ellipsoided in a diagonal cross section on the whole body and 2) incessant wiggly movement pattern as that of fly maggot, with the larvae of the above mentioned species. These characters seem to reflect the phylogenetic relationships among acheilognathine fishes. On the other hand, this species and Acan. asmussi are spring-summer spawning bitterlings. And also these species never retard the larval growth in such larval stage as the duration from Stage B to Stage D.