Rhodeus Ocellatus Kurumeus Research Articles

A ddRAD-based population genetics and phylogenetics of an endangered freshwater fish from Japan

Rhodeus ocellatus kurumeus is a small cyprinid fish endemic to the Japanese archipelago. This fish frequently hybridizes with an exotic subspecies Rhodeus ocellatus ocellatus, and non-introgressed populations of R. o. kurumeus have greatly decreased. Previous studies based on mitochondrial and nuclear DNA identified introgressed populations and inferred phylogenetic relationships, but these approaches may lead to underestimates of introgression or ambiguous results owing to the small number of genetic markers used. The present study applied double digest restriction site-associated DNA (ddRAD) sequencing to assignment tests and phylogenetic inferences. Data matrices can vary in size and content depending on the strategies used to process ddRAD sequences; therefore, we prepared 25 data matrices with various processing strategies. The assignment tests based on the 25 data matrices resulted in similar assignment patterns and provided evidence for introgression in some populations that had been identified as non-introgressed populations in previous studies. The maximum-likelihood (ML) phylogenetic trees differed in robustness and topology among the 25 matrices; interestingly, ML trees with low statistical support reflected geographical distributions of fish better than ML trees with high statistical support. These results indicate that ddRAD sequencing can detect introgression with greater sensitivity than conventional DNA markers, that ddRAD sequencing is useful for phylogenetic inference among closely related populations within a subspecies, and that statistically robust ML trees do not necessarily reflect the true phylogeny. Application of ddRAD data to conservation genetics and evolutionary history of this subspecies are also discussed.

マイクロサテライトマーカーから見た大阪産ニッポンバラタナゴ溜池集団の遺伝的特徴

溜池に生息する大阪産ニッポンバラタナゴについてマイクロサテライトマーカーを用いた遺伝的特徴の推定を行った。平均アリル数は何れの集団も約 2 と低い値を示したが，ヘテロ接合度と血縁度は集団間で大きく異なり，その要因として創始者効果と生息地の規模の違いが考えられた。また，アリル組成において集団間の違いは殆ど見られなかったが，南北間で遺伝的分化が認められた。これらの結果は大阪集団が単一起源である一方，ボトルネックにより遺伝的分化が生じており，近親交配の回避において環境収容力が重要であることを意味する。

An extant native population of the Japanese rosy bitterling, Rhodeus ocellatus kurumeus, in the Sanyo Region, western Japan

An extant native population of the Japanese rosy bitterling, <i>Rhodeus ocellatus kurumeus</i>, in the Sanyo Region, western Japan

Predicting distributions of seven bitterling fishes in northern Kyushu, Japan

The distributions of seven bitterling species and subspecies—Tanakia lanceolata, T. limbata, Acheilognathus tabira nakamurae, A. rhombeus, Rhodeus ocellatus kurumeus, R. ocellatus ocellatus, and R. atremius atremius—in northern Kyushu were predicted using generalized linear models (GLMs) in order to provide information helpful for conserving native bitterlings and preventing the expansion of alien bitterling species. Predictions were made according to the following procedure: (1) a set of GLMs for each species was formulated using environmental data from 710 sites that were derived using digital maps and GIS software, from which the best fit model for each species was selected using the Akaike information criterion for predicting the fish occurrence, (2) model performance was evaluated based on the receiver-operating characteristics (ROC) analysis using occurrence and environmental data from 362 sites, and (3) potential distributions of the bitterling were analyzed using the best fit models and environmental data for 1,272 sites, of which 200 data points without occurrence data were prepared. The best fit models revealed that 4–6 environmental factors were important in predicting seven bitterling distributions, which was supported by the area under the ROC curve (AUC) values of these fishes ranging from 0.753 to 0.927. The AUC values in model evaluation were significantly greater than 0.5 for six fishes, suggesting the moderate accuracies of these best fit models for predicting the fish distributions. These predictive models can be used for evaluating potential native bitterling richness and the potential distribution expansion of an alien subspecies.

Discovery of the Japanese rosy bitterling, Rhodeus ocellatus kurumeus, in Nara Prefecture

Discovery of the Japanese rosy bitterling, Rhodeus ocellatus kurumeus, in Nara Prefecture

Isolation and characterization of new microsatellite markers for rose bitterlings, Rhodeus ocellatus

The Japanese rose bitterling (Rhodeus ocellatus kurumeus) is facing imminent extinction because of hybridization and competition from an invasive alien subspecies (Rhodeus ocellatus ocellatus). Eleven new microsatellite markers for the two subspecies were developed using dinucleotide repeat specific polymerase chain reaction. The number of alleles per locus and the heterozygosity in R.o. kurumeus were lower than those in R.o. ocellatus. Most of these microsatellite markers were successfully cross-amplified in three Acheilognathinae species.

希少淡水魚ニッポンバラタナゴ保護の取組み―タイリクバラタナゴ混入個体群の池干しと捕獲による防除の試み―

Japanese rose bitterling, Rhodeus ocellatus kurumeus, is an endangered freshwater fish in Japan. Both the expansion of the habitat of an alien subspecies, R. o. ocellatus, and the increased chances of hybridization between the two subspecies have threatened the genetic identity of R. o. kurumeus. To conserve R. o. kurumeus in Kagawa Prefecture, 17 populations were genetically monitored in 2006. Two individuals in a pond were found to harbor R. o. ocellatus type mitochondrial DNA through PCR-RFLP analysis. Then the contaminated population was transplanted to a closed Bio-top water system to prevent contamination in other ponds. Three-day pond drying was found to be insufficient to eradicate R. ocellatus.

Timing of juvenile emergence from host mussels in the Japanese rosy bitterling, Rhodeus ocellatus kurumeus

The diel rhythm of juvenile emergence from host mussels in the Japanese rosy bitterling, Rhodeus ocellatus kurumeus, was investigated in a small pond in Osaka prefecture, Japan. The emergence of juvenile bitterling from host mussels occurred principally at night, with a peak in the early part of the night. The pattern of juvenile bitterling emergence was discussed in the context of an adaptive response to diurnal predation.

有明海沿岸域のクリークにおける淡水魚類の生息の有無・生息密度とクリークの護岸形状との関係

The presence and populations of freshwater fishes and the revetment conditions in creeks were investigated at 48 sampling sites around the Sea of Ariake in northwestern Kyushu, Japan. Each creek studied in this survey was classified into one of the three following types in terms of revetment conditions: Type-1, the creeks have earthen revetments except when crossing roads; Type-2, although there are concrete revetments in the creeks, these revetments are short, and agricultural water flows over the revetments and reaches bank vegetations or earthen banks except during the agricultural off-season; Type-3, there are high concrete revetments around the creeks, and agricultural water does not flow over the revetments during any season. The amount of Hemigrammocypris rasborella present was affected by the revetment conditions, and the amount present in Type-3 was significantly lower than those in Type-1 and Type-2. The populations of H. rasborella, Rhodeus ocellatus kurumeus, and Pseudorasbora parva were affected by the revetment conditions. For H. rasborella and R. ocellatus kurumeus, the populations in Type-3 were significantly lower than those in Type-1 and Type-2. For P. parva, the population in Type-3 was significantly lower than that in Type-1.

Adaptive spatial utilization of host mussels by the Japanese rosy bitterling Rhodeus ocellatus kurumeus

The spatial pattern of resource utilization for oviposition in the Japanese rosy bitterling Rhodeus ocellatus kurumeus was investigated in two field experiments. The distribution of rosy bitterling eggs deposited in the four demibranchs of the gills of the test mussel species, Anodonta woodiana, differed with mussel reproductive state in pair spawnings, but not in group spawnings. In pair spawnings, female rosy bitterling may have had more time to select the site of oviposition in the gill in relation to the sex and reproductive state of the mussel, thereby maximizing embryo survival. Thus, the inner gill of female mussels may have been selected in preference to the outer gills to avoid mortalities of eggs due to the presence of the mussel's own embryos (glochidia) in the outer gill chambers. In male mussels, female rosy bitterling distributed their eggs equally among all parts of the mussel gill, thereby minimizing density‐dependent mortality of embryos. During group spawnings, however, female rosy bitterling may have been more constrained in their decision making, ovipositing in the inner gills irrespective of mussel sex.

Seasonal change in the spatial utilization of host mussels in relation to ovipositor length by female rosy bitterling Rhodeus ocellatus kurumeus

The seasonal changes in ovipositor length and utilization patterns of mussels for oviposition in the rosy bitterling Rhodeus ocellatus kurumeus(Cyprinidae) were investigated in a field experiment and field surveys during the breeding period (April to August). The mean length of ovipositors at oviposition was short at the start (early April) and end (July) of the breeding period. Females with long ovipositor at oviposition were collected between mid‐April and June. Mark‐and‐recapture data showed that ovipositor length at oviposition changed rhythmically throughout the breeding period, shortening and lengthening as the female entered the spawning and resting phase. The density of rosy bitterling embryos in mussels increased between April and June, peaking in May, but decreasing in July. The position of eggs on mussel gills varied from close to the exhalant siphon to deeper inside the gill during April, and periodically thereafter. There was a positive correlation between ovipositor length at oviposition and the distance from exhalant siphon of mussels to eggs deposited by females, suggesting that ovipositor length at oviposition determined the position of eggs deposited on a mussel gill. Because dissolved oxygen in mussel gills decreased with the density of bitterling embryos, suitable positions for embryo survival in gills changed with embryo density. By changing ovipositor length at oviposition, females might be able to spawn their eggs in a position that maximizes embryo survival. Thus, plasticity in ovipositor length at oviposition may play an important role as an adaptation of rosy bitterling in utilizing mussels when their quality as a spawning substratum fluctuates seasonally.

多々良川水系におけるタナゴ類の分布域の推移とタナゴ類・二枚貝の生息に及ぼす都市化の影響

The populations of Japanese bitterlings and unionid mussels and the land use of the watershed were investigated in 36 sampling sites in the Tatara river system in northern Kyushu, Japan. Five bitterling species were found in 11 sites in the system. Although Rhodeus ocellatus kurumeus was found to be distributed in 23 sites in 1983, this species was found in three sites in this study. The population of the bitterlings decreased in the sites with a high urbanization rate, although the populations of several other fish species showed no dependence on the urbanization rate. The population of the mussels showed a negative correlation with urbanization rate. In addition, the mussels populations showed positive relationship with the bitterling populations. These three relationships indicate that the decrease in the bitterling population due to the urbanization of the watershed was responsible for the decrease in the mussel population.

Factors affecting seasonal mortality of rosy bitterling (Rhodeus ocellatus kurumeus) embryos on the gills of their host mussel

Factors affecting seasonal mortality of rosy bitterling (<i>Rhodeus ocellatus kurumeus</i>) embryos on the gills of their host mussel

Low Genetic Variation and Inbreeding Depression in Small Isolated Populations of the Japanese Rosy Bitterling, Rhodeus ocellatus kurumeus

The Japanese rosy bitterling, Rhodeus ocellatus kurumeus, has been affected not only by the invasion of another subspecies, R. o. ocellatus, from China, but also by habitat fragmentation. In this study, the effects of habitat fragmentation on the fitness of R. o. kurumeus were investigated. Owing to exclusion by R. o. ocellatus, R. o. kurumeus in Honshu and Shikoku has disappeared entirely, except for small populations in isolated man-made ponds in Osaka and Kagawa. In Kyushu it still occupies open water systems, into which R. o. ocellatus has only recently invaded. Meristic and genetic data show that the diversity of R. o. kurumeus is significantly lower in the isolated Osaka and Kagawa populations than the non-isolated Fukuoka population. The Osaka population is inferior to the Fukuoka population in terms of viability and growth. The viability of reciprocal inter-population hybrids between the Osaka and Fukuoka populations was, however, as high as that of the Fukuoka population. In addition to the high scores of band sharing index (BSI) in RAPD-PCR analysis, acceptance of transplanted scales among individuals, irrespective of natal pond, indicates that the Osaka population forms a highly inbred line. These results suggest that low genetic variation is associated with inbreeding depression in the small isolated Osaka populations. Consequently, the management of ponds, including the free movement of individuals, in addition to measures to prevent the invasion of R. o. ocellatus, is necessary for the conservation of R. o. kurumeus.

Factors affecting seasonal mortality of rosy bitterling (Rhodeus ocellatus kurumeus) embryos on the gills of their host mussel

AbstractI investigated the seasonal change in factors affecting embryonic mortality in the rosy bitterling, Rhodeus ocellatus kurumeus, a freshwater fish that spawns on the gills of living unionid mussels. Research was conducted in a small pond during 1999 and 2001 in which bitterling were provided with Anodonta sp. mussels for spawning. Bitterling spawned between April and July, peaking mid–late May. Seasonal survival rate of bitterling embryos in their mussel hosts was unimodal, with a peak between late April and mid May (about 70% of total spawnings). In mid April, survival was about 50%. The lowest survival was from late May to July (0%). Losses of bitterling embryos from mussels were identified by ejections from the mussel host. Ejections were categorized as either ejections of live embryos, or ejections of embryos that died in the mussel and were subsequently expelled from the mussel. Ejection rates of live embryos were higher in the earlier part of the spawning period (early–mid April) and dead embryo ejections in the later period (after June). The ejection rate of live embryos was higher among younger embryos earlier in the season, probably because of the incomplete development of morphological and behavioural traits associated with maintaining the embryo inside the mussel gill chambers, and as a consequence of a more protracted developmental period at low temperatures making them more susceptible to ejection. The ejection rate of dead embryos was higher in older embryos later in the season, and in larger mussels and at high embryo densities. The survival of embryos in mussels was probably related to oxygen availability, with mortalities probably caused by asphyxiation. Increased embryo mortalities may arise through competition among embryos, between embryos and mussel, and ambient dissolved oxygen levels. The optimal period for bitterling to spawn may represent a balance between two opposing factors; with positive and negative effects of a seasonal rise in temperature directly affecting embryonic growth rate and oxygen availability.

FISH-mapping of 18S ribosomal RNA genes and telomeric sequences in the Japanese bitterlings Rhodeus ocellatus kurumeus and Tanakia limbata (Pisces, Cyprinidae) reveals significant cytogenetic differences in morphologically similar karyotypes.

The Japanese rose bitterling, Rhodeus ocellatus kurumeus, and the oily bitterling, Tanakia limbata, were cytogenetically studied by silver (Ag)- and chromomycin A3 (CMA3)-staining, by C-banding and by mapping of the 18S ribosomal genes and of the (TTAGGG)n telomeric sequence. These two representative species of related genera of the subfamily Acheilognathinae show very similar chromosome complements. Nevertheless, significant differences in the chromosomal distribution of nucleolus organizer regions (NORs) and interstitial telomeric sequences were observed. Whereas R. ocellatus kurumeus shows a single NOR-bearing chromosome pair, T. limbata is characterized by a higher number of variable NORs. Multiple telomeric sequence sites were found at the pericentromeric regions of several chromosomes in the rose bitterling. No telomeric sequence sites were detected near centromeres, but they were found to be scattered along the NORs in the oily bitterling. Two karyoevolutive trends might have been identified in the subfamily.

Filial cannibalism improves survival and development of beaugregory damselfish embryos

Cannibalism of small numbers of offspring by a parent has been proposed as an adaptive parental strategy, by providing energy to support parental care. However, there are few empirical studies to support this hypothesis. We conducted field and laboratory experiments to investigate partial filial cannibalism in Stegastes leucostictus, a coral reef fish with paternal care. Partial cannibalism was shown to be common, and males were found to remove developing embryos from throughout a clutch in a random pattern, rather than in the more aggregated pattern seen during embryo predation. Males that received a diet supplement grew faster than control males, but did not engage in less cannibalism. Also, males did not concentrate cannibalism on early embryonic stages with the highest energetic value. Experimental reduction of embryo densities was found to significantly increase embryo development rate and survival from egg deposition to hatching, and experimental reduction of oxygen levels significantly increased rates of partial filial cannibalism by males. Artificial spawning sites with low oxygen levels were avoided by spawning females, and cannibalism rates by males were higher. We propose that partial filial cannibalism serves as an adaptive parental strategy to low oxygen levels in S. leucostictus by increasing the hatching success of embryos.

Genetic diversity in the Japanese rosy bitterling, Rhodeus ocellatus kurumeus (Cyprinidae)

Comparison of meristic characters (pored lateral line scales, vertebrae, and fin rays), and PCR-RFLP analysis in the D-loop and ND1 regions of mitochondrial DNA were performed to estimate the genetic diversity in local populations of the Japanese rosy bitterling, Rhodeus ocellatus kurumeus. In terms of meristic characters, the Fukuoka population was the largest in both range and variance of the number of pored lateral line scales and vertebrae (abdominal and caudal), and Osaka was the second, whereas the Kagawa population showed the smallest range and variance in these characters. In PCR-RFLP analysis, 11 haplotypes (3 in Fukuoka, 2 in Okayama, 2 in Kagawa, and 4 in Osaka) were observed, and nucleotide sequence divergence (NSD) was approximately two times larger in ND1 (mean, 0.61%) than in D-loop (mean, 0.31%). In the neighbor-joining (NJ) tree, based upon the NSD value in ND1, haplotypes were arranged into four clades, which corresponded to the locality of each haplotype. The Fukuoka population was conspicuously apart from the other populations (mean, 0.90% in NSD), but the remaining three showed a similar genetic distance with each other (mean, 0.48%–0.52% in NSD). In haplotype diversity of mtDNA, half the stations in Osaka and all in Kagawa were monomorphic. Especially, two haplotypes endemic to Kagawa were randomly distributed, irrespective of drainages. Rhodeus o. kurumeus in Fukuoka inhabits small rivers and creeks (open water systems), while that in Kagawa and Osaka lives in small ponds (closed water systems). Taking the information of morphology, mtDNA, and habitat into consideration, the low genetic diversity in Kagawa and Osaka populations of R. o. kurumeus is thought to be mainly the result of the isolation of their habitat.

Genetic Introgression by the Rose Bitterling, Rhodeus ocellatus ocellatus, into the Japanese Rose Bitterling, R. o. kurumeus (Teleostei: Cyprinidae)

The Japanese rose bitterling, Rhodeus ocellatus kurumeus, is an endangered cyprinid species. The main reason of drastic diminution of R. o. kurumeus has been suggested to be due to hybridization between this subspecies and R. o. ocellatus introduced from China. Both RFLP analysis of mitochondrial DNA and RAPD-PCR analysis of genomic DNA were performed to elucidate the genetic introgression of R. o. ocellatus into R. o. kurumeus. The two subspecies were distinguished in the D-loop region of mtDNA by six restriction endonucleases. Rhodeus o. kurumeus had eleven subspecies-specific RAPD markers and R. o. ocellatus had two. Except some R. o. kurumeus populations, most of the populations of R. ocellatus in Japan were hybrids, equipped with subspecies-specific RAPD markers for the two subspecies, respectively. The genetic constitution of these markers in hybrids, however, greatly differed among populations. The R. o. ocellatus mtDNA was predominantly observed in hybrid populations, except two populations with mtDNAs of the two subspecies. Judging from the genetic dominance of morphological and physiological characters of R. o. ocellatus against R. o. kurumeus, hybrids probably have the same ecological dominance as R. o. ocellatus against R. o. kurumeus. Therefore, it is considered that R. o. kurumeus not only has its genetic property spoiled by hybridization with R. o. ocellatus, but also is expelled by R. o. ocellatus and hybrids. The replacement of mtDNA and genomes of R. o. kurumeus with those of R. o. ocellatus in hybridization might be accelerated by the backcross between hybrids and R. o. ocellatus.