Host size matters for reproduction: Evolution of spawning preference and female reproductive phenotypes in mussel-symbiotic freshwater bitterling fishes.

Bitterling fishes evolve an idiosyncratic symbiosis with freshwater mussels, in which they are obligated to spawn in the gills of mussels for reproduction. In recent years, freshwater mussel populations have been drastically diminishing, due to accelerating anthropogenic impacts, which can be large threats to the risk of bitterling's extinction cascade (i.e. 'coextinction'). The host mussel size may be an important factor driving the adaptation and evolution of bitterling's reproductive phenotypes. Here we examined the host size preference and morphological adaptation of female bitterling to the host size from 17 localities at the Han River in Korea. Using our developed molecular-based species identification for bitterling's eggs/larvae inside the mussels, we further determined the spawning patterns of seven bitterling species. Mean length of spawned mussels (N = 453) was significantly larger than that of unspawned mussels (N = 1814), suggesting that bitterling prefers to use larger hosts as a spawning ground. Spawning probability was clearly greater as mussel size increases. Results of our reciprocal transplant experiments do provide some evidence supporting the 'bitterling's larger host preference' hypothesis. Interspecific competition appeared to be intense as two fish species often spawned eggs in the same mussel individuals simultaneously. Longer ovipositor and more elongated egg may evolve in females of Tanakia signifer in response to larger host environments. The observed bitterling's spawning preference for large-sized mussels may evolve perhaps because of the fitness advantage in relation to the offspring survival. Our findings further inform on the development of effective conservation and management strategy for the endangered bitterling fishes.

On the Species Identification of Two Non-Native Tilapia Species, Including the First Record of a Feral Population of Oreochromis aureus (Steindachner, 1864) in South Korea.

Tilapia is an invasive species that has become widely distributed around the world. In Korea, introduced tilapia into its aquatic ecosystem for the first time with a species from Thailand in 1955, and later additionally introduced two more species from Japan and Taiwan, thus securing a total of three species of tilapia (O. niloticus, O. mossambicus and O. aureus) as food resources. Since then, O. niloticus has been reported to inhabit certain streams with thermal effluent outlets. Morphological species identification is very difficult for tilapia and a combined analysis of morphological and molecular-based species identification is therefore necessary. This study investigated a tilapia population that inhabits a thermal effluent stream (Dalseo Stream) in Daegu Metropolitan City, Korea, in order to conduct a morphological and genetic species identification of this population. In total, 37 tilapia individuals were sampled. The results of the morphological and genetic species identification analyses found that two species, O. aureus and O. niloticus, inhabit the Dalseo Stream. In Korea, the habitat of the O. niloticus natural population has been reported, but the O. aureus natural population has not been reported. Thus, we observed for the first time that a new invasive species, O. aureus, inhabits a stream in Korea. They are known to cause disturbances to aquatic organisms (e.g., fish, aquatic insects, plankton, aquatic plants) and the habitat environment (e.g., water quality, bottom structure). Accordingly, it is important to study the ecological effects of O. aureus and O. niloticus on the corresponding freshwater ecosystem closely and to prepare a management plan to prevent the spread of these species, as they are notoriously invasive.

Complete mitochondrial genome of a Korean endemic freshwater mussel Nodularia breviconcha (Bivalvia: Unionidae).

Mitochondrial genome sequences were first determined and analyzed for a Korean endemic freshwater mussel Nodularia breviconcha (synonym Nodularia douglasiae sinuolatus; Unionidae, Unionida, Bivalvia). The complete mitochondrial genome was 15,741 bp in length, including 13 protein-coding genes (PCGs), 22 tRNA genes, and 2 rRNA genes. The overall GC content of mitochondrial genome for N. breviconcha was 34.3%. Phylogenetic analysis of 18 species within the family Unionidae suggested that Nodularia douglasiae is the most closely related to N. breviconcha. Our study will provide baseline, but important information for future research on ecological and genetic/genomic characteristics of this species.

Assessment of fish biodiversity in four Korean rivers using environmental DNA metabarcoding.

Environmental DNA (eDNA) metabarcoding is a cost-effective novel approach to estimate biodiversity in an ecosystem. In this study, the MiFish pipeline was employed to test if the system methodology is sufficiently reliable to estimate fish biodiversity in Korean rivers. A total of 125 unique haplotypes and 73 species were identified at the species level from 16 water samples collected from a single survey in four Korean rivers (Hyeongsan, Taehwa, Seomjin, and Nakdong). Among the four rivers, the highest species richness was recorded in the Seomjin River (52 species), followed by the Taehwa (42 species) and Hyeongsan (40 species) rivers. The Nakdong River (26 species) presented the lowest species richness and number of endemic species, presumably due to its metropolitan location and anthropogenic impacts, such as dams or weirs. We were also able to detect that five exotic species (Carassius cuvieri, Cyprinus carpio, Cyprinus megalophthalmus, Lepomis macrochirus, and Micropterus salmoides) are widely distributed in all surveyed rivers, a situation that might be problematic in terms of conservation. Our findings indicate that the eDNA metabarcoding technique is one of the most cost-effective scientific tools available for the management and conservation of the freshwater fish resources available in Korea. However, the low number of 12S sequences of endemic species in the database and low resolution of the MiFish region for differentiating several taxa should be upgraded for their wide use.