Masou Research Articles

Masu salmon species complex relationships and sex chromosomes revealed from analyses of the masu salmon (Oncorhynchus masou masou) genome assembly.

Masu salmon (Oncorhynchus masou) are the only Pacific salmon endemic to Asia. Some researchers prefer to categorize these salmon into four subspecies (masu - O. m. masou, amago - O. m. ishikawae, Biwa - O. m. subsp., and Formosan - O. m. formosanus), while others prefer individual species designations. Even though the masu salmon fishery is thousands of years old, classification of the diversity within the masu salmon species complex remains elusive. In this study, a genetic map and reference genome assembly were generated for one species/subspecies (masu) to provide resources for understanding the species complex. In O. m. masou, the sex chromosome was determined to be chromosome 7. Resequenced genomes from two other putative subspecies (amago and Biwa) provided evidence that they do not share the same sex chromosome. Principal component and admixture analyses clustered the amago and Biwa salmon close together. This supported previous findings of a close relationship between amago and Biwa salmon and a more distant relationship to masu salmon for both. Additional analyses of the masu salmon species complex will benefit from using the new reference genome assembly.

Distinct food-web transfers of 137Cs to fish in river and lake ecosystems: A case study focusing on masu salmon in the Fukushima evacuation zone

This study was conducted to elucidate the spatial and size variations, and food-web transfer of 137Cs in freshwater fish in the upper reaches of the Ukedo River system, a highly contaminated river system flowing through the Fukushima evacuation zone. Fish collection and environmental surveys were conducted in the summer of 2020 at five forest rivers and at the Ogaki Dam reservoir (an artificial lake) with different air dose rates (mean 0.20–3.32 μSv/h). From the river sites, two salmonid species (masu salmon and white-spotted charr) were sampled, with masu salmon generally exhibiting higher 137Cs concentrations, ranging widely (10.6 Bq/kg-wet to 13.0 kBq/kg-wet) depending on the fish size (size effect) and site. The 137Cs concentrations in masu salmon were explained by the air dose rates, 137Cs concentrations in water, sediments (excluding the lake site), and primary producers, with site-specific variations. In the rivers, masu salmon (fluvial type with parr marks) mainly fed on terrestrial insects with higher 137Cs concentrations compared with those of aquatic insects, indicating that 137Cs was transferred mainly to fish through the allochthonous forest food-web during summer. In the lake, masu salmon (lake-run type with larger size and silvery body coloration) mainly preyed on smaller fish with lower 137Cs concentrations, demonstrating that 137Cs is transferred to fish through the autochthonous lake food-web with biomagnification. Differences in 137Cs concentrations among masu salmon (mean 441 Bq/kg-wet) and other fish species (mean 74.8 Bq/kg-wet to 2.35 kBq/kg-wet) were also found in the lake. The distinct 137Cs transfers to river and lake fish were supported by stable isotope analysis: δ15N and δ13C values enriched stepwisely through the food-webs were, respectively, higher and lower in the lake. Our results obtained using multiple approaches clearly revealed the distinct food-web transfer of 137Cs in river and lake ecosystems. These findings can contribute to prediction of radioactive contamination in freshwater fish in the Fukushima evacuation zone.

Postharvest quality evaluation of masu salmon (Oncorhynchus masou) during ice storage by spinal cord and bleeding

The study investigated the impact of spinal cord-treated and bleeding processes on the quality changes of postharvest aquaculture masu salmon during ice storage. Fish underwent ice storage for 0–70 h, and the rigor mortis, ATP-related compounds, K-value, free amino acids, drip loss, and pH were monitored. Spinal cord-treated and bleeding fish started rigor mortis at 1.5 h, reaching full at 4.5 h, and persisted for 21 h, while control samples exhibited an earlier onset of rigor mortis at 0.5 h, reaching a maximum of 1 h. The ATP content showed a significant difference during the initial storage period from 0 to 3 h, with spinal cord-treated and bleeding fish showing a higher ATP content and lower K-value, and followed a similar trend, reaching around 20–23 % after 70 h of storage. The most abundant free amino acid detected was Ans, which occupied over 70 %, with others including Tau, Gln, Thr, Ser, Gly, Ala, His, and Glu. Spinal cord-treated and bleeding fish exhibited higher contents of Thr, Ser, and Gly, contributing to sweetness (p < 0.05). The umami flavor compound Glu increased with storage time, particularly in the spinal cord and bleeding fish, while IMP was most abundant during the mid-storage period, with higher levels in the spinal group with the prolong of storage time. HxR and Hx primarily contributed to bitterness, increasing with storage time, whereas His did not show a significant increase in the later storage period. Spinal cord and bleeding treatment demonstrated reduced drip loss (p < 0.05). The pH value fluctuated between 6.2 and 6.5 with no significant differences between the two treatments. The spinal cord-treated and bleeding treatments minimized stress and resulted in overall better postharvest quality, aligning with fish welfare.

Morphological and Molecular Characterization of Three Myxosporean Species of the Genera Myxobolus, Henneguya, and Myxidium (Cnidaria: Myxozoa) Infecting Freshwater Fish, Isolated for the First Time in Japan.

The majority of myxosporean species (Cnidaria: Myxozoa) of the genera Myxobolus (35 species), Henneguya (8 species), and Myxidium (9 species) from freshwater or brackish fish in Japan were recorded more than 30 years ago (accumulatively 81.1% [43/53]). The re-discovery and molecular-genetic characterization of these species is a current research priority. During our myxosporean survey in Japanese freshwater fish, we detected three species that had never been recorded in Japan, but in the Russian Far East (Sakhalin Island, and Maritime Province): Myxobolus tribolodonus sp. n., forming cysts in the gills of Tribolodon sachalinensis (syn. M. marinus sensu Aseeva, 2000; M. marinus sensu Sokolov et Frolova, 2015, recorded from the gills of Pseudaspius (syn. Tribolodon) spp.); Henneguya pungitii Achmerov, 1953, forming cysts in the subcutis of external skin and buccal submucosa of Pungitius sinensis; and Myxidium salvelini Konovalov et Shulman, 1966, in the urinary bladder of Oncorhynchus masou ishikawae. These new isolates were characterized by integrated taxonomic approaches, i.e., myxospore morphology and molecular-genetic characterization of the small subunit ribosomal RNA gene (SSU rDNA). These new isolates were phylogenetically differentiated from any species whose SSU rDNA sequences were deposited in the DNA databases, and concurrently compared with recorded species based on classical morphological criteria. All three species were differentiated from myxosporeans previously recorded in Japan, indicating new distribution records out of the Russian Far East. For reliable species identification, accumulation of at least SSU rDNA sequences of known species worldwide is critically important.

Messenger RNA transcription levels of neuronal soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex components in the olfactory nerve system of the anadromous Pacific salmon, masu salmon Oncorhynchus masou.

Anadromous Pacific salmon (genus Oncorhynchus) are known for homing behavior to their natal rivers based on olfactory imprinted memories during seaward migration. The SNARE complex is a regulator of vesicle exocytosis from the presynaptic membrane. Our previous study suggested that its component genes (Snap25, Stx1, and Vamp2) are more highly expressed in the olfactory nervous system (ONS) during the migration stages associated with olfactory imprinting in the evolutionary species of Pacific salmon, such as chum (O. keta) and pink (O. gorbuscha) salmon. Masu salmon (O. masou) has a significantly different life history from these species, living longer in rivers and being a more primitive Pacific salmon species. In this study, the transcription of snare mRNAs in the ONS was analyzed using mainly male wild masu salmon. Five cDNAs encoding masu salmon SNAREs, which are well conserved among vertebrates, were isolated and sequenced. Each snare mRNA was highly expressed in age 1+ (yearling) parr prior to smoltification, particularly in the olfactory bulb. Their transcription status was significantly different from that of chum and pink salmon, which showed high expression in earlier under-yearling juveniles. The present results and our previous studies indicate that snare mRNAs are highly transcripted until the seaward migration, reflecting neural development and neuroplasticity of the ONS for olfactory imprinting.

A comparison of sex‐specific markers for two wild masu salmon populations in Hokkaido, Japan

AbstractWe evaluated the utility of three male‐specific molecular markers, sdY, sdY 227U and OtY2m, in two wild populations of masu salmon (Oncorhynchus masou) in southwestern Hokkaido, Japan. Male‐specific fragments amplified in all phenotypic males across the markers. Phenotypic females were genetically identified as males using sdY (57%) and sdY 227U (4%), but no phenotypic females were identified as males using OtY2m. We conclude that OtY2m was the most reliable sex marker, followed closely by sdY 227U, among those tested in our study populations. Additional research is warranted to test the applicability of these markers in other populations and Oncorhynchus species.

Optimizing the post-thaw quality of cryopreserved masu salmon (Oncorhynchus masou) sperm: Evaluating the effects of antioxidant-supplemented extender

The oxidative damage induced by cryopreservation significantly contributes to the overall decline in the quality of cryopreserved sperm. This underscores the necessity for exploring mitigation measures, such as supplementing the freezing media with antioxidants, aimed at enhancing sperm quality post-thaw. In this study, we determined the effects of adding five antioxidants [Vitamin C, taurine, trolox, Butylhydroxytoluene (BHT), and uric acid] to a glucose–methanol extender at three concentrations—low, intermediate, and high—on the motility, viability, concentration, and fertilization rates of the sperm of masu salmon (Oncorhynchus masou), which is an economically important fish resource in Northeast Asia. Our research reveals various sperm quality parameters of the same species react differently to different antioxidants or even different concentrations of the same antioxidants, which adds another layer of complexity to the selection criteria for an appropriate antioxidant for supplementation beyond the specific specificity of antioxidants. Moreover, the effects of the antioxidants were often nonlinear and paradoxical, highlighting the importance of applying accurate dosage. Compared with the control group (no antioxidant supplementation), only uric acid and taurine supplementation at intermediate concentrations (5 mM) significantly improved the post-thaw sperm viability and percentage of motile sperm. Meanwhile, BHT supplementation generally improved the following kinematic traits of sperm: straight-line velocity (VSL), linearity, average path velocity, and curvilinear velocity (VCL). Similarly, 5-mM taurine supplementation also significantly improved sperm VSL, VCL, and beat cross frequency. In contrast, 10-mM Vitamin C supplementation (high concentration) adversely affected multiple parameters, leading to a decline in fertilization capacity. Based on these findings, the selection criteria for antioxidants should go beyond species specificity and include sperm quality parameters the breeder considers crucial for predicting post-thaw sperm fertility, as well as the ways in which particular antioxidants influence these parameters. We recommend 5-mM taurine supplementation for masu salmon sperm cryopreservation because of its positive effects on sperm viability, concentration, and some kinematic parameters.

Correction: Exploring predators of Pacific salmon throughout their life history: the case of Japanese chum, pink, and masu salmon

Correction: Exploring predators of Pacific salmon throughout their life history: the case of Japanese chum, pink, and masu salmon

Optimal SNP filtering strategies for pedigree reconstruction: A case study with wild red‐spotted masu salmon population

Optimal <scp>SNP</scp> filtering strategies for pedigree reconstruction: A case study with wild red‐spotted masu salmon population

Some features of natural spawning and problems of artificial reproduction of the cherry salmon Oncorhynchus masou (Brevoort, 1856)

The article discusses certain aspects of the dispersal and biology of cherry salmon, Oncorhynchus masou (Brevoort, 1856). It is noted that the species is the southernmost within the Pacific Salmon family and is also the only member of the Oncorhynchus genus whose habitat is exclusively in Asia. The decline in the number of cherry salmon across their range has been highlighted, and the main underlying factors have been identified. Some features of cherry salmon spawning run in various parts of their range are examined. A long stay of spawners in fresh water before spawning with preservation of feeding behavior is noted, which makes the species an extremely compelling object of recreational and commercial fishing. Some features of natural reproduction, such as spawning timing and fertility, depending on the habitat and size of the spawners, are indicated. The issue of preserving and restoring the species population is touched upon; the most appropriate method of its implementation is the artificial reproduction of the young with subsequent release into the natural habitat. The paper highlights a number of obstacles to the implementation of such programs, such as the undeveloped technology for growing young masu salmon and the difficulty of optimizing the temperature regime at hatcheries. As one of the limitations to increasing release volumes, it is indicated that masu salmon belongs to a species that have a long freshwater period, with the young spending up to three years in the river. A large number of juvenile masu salmon in the river leads to insufficient food supply and high mortality. In order to overcome this factor, it has been proposed to conduct research aimed at stimulating the smoltification of hatchery salmon fingerlings. This measure will allow significantly increasing the number of smolt and the return rates of spawners, which can be removed without affecting the natural populations.

The upper limit of redd abundance compared to the numbers of anadromous adult masu salmon in Horokashubuto Stream, Hokkaido, northern Japan

The upper limit of redd abundance compared to the numbers of anadromous adult masu salmon in Horokashubuto Stream, Hokkaido, northern Japan

Comparative analysis of ventricular stiffness across species.

Investigating ventricular diastolic properties is crucial for understanding the physiological cardiac functions in organisms and unraveling the pathological mechanisms of cardiovascular disorders. Ventricular stiffness, a fundamental parameter that defines ventricular diastolic functions in chordates, is typically analyzed using the end-diastolic pressure-volume relationship (EDPVR). However, comparing ventricular stiffness accurately across chambers of varying maximum volume capacities has been a long-standing challenge. As one of the solutions to this problem, we propose calculating a relative ventricular stiffness index by applying an exponential approximation formula to the EDPVR plot data of the relationship between ventricular pressure and values of normalized ventricular volume by the ventricular weight. This article reviews the potential, utility, and limitations of using normalized EDPVR analysis in recent studies. Herein, we measured and ranked ventricular stiffness in differently sized and shaped chambers using exvivo ventricular pressure-volume analysis data from four animals: Wistar rats, red-eared slider turtles, masu salmon, and cherry salmon. Furthermore, we have discussed the mechanical effects of intracellular and extracellular viscoelastic components, Titin (Connectin) filaments, collagens, physiological sarcomere length, and other factors that govern ventricular stiffness. Our review provides insights into the comparison of ventricular stiffness in different-sized ventricles between heterologous and homologous species, including non-model organisms.

Genetic characterization of juvenile chum salmon with otolith marks of Japanese salmon hatcheries in the catches of trawl surveys 2016–2021

Chum salmon sample from the catches of autumn trawl surveys in the Sea of Okhotsk (2016–2021), including individuals with otolith marks of Japanese salmon hatcheries (SHs), was analyzed based on microsatellite loci polymorphism. Reference database of eight microsatellite loci frequencies, created before and included 26 samples of chum salmon from major reproductive regions in the Sea of Okhotsk basin, has been updated with data on the microsatellite allele variety of chum salmon from Hokkaido and Honshu. Result s of the research indicate of strong similarity between chum salmon of the Kuril Islands and individuals released from Japanese SHs.

The strength of density dependence on body size of young‐of‐the‐year masu salmon, Oncorhynchus masou, varies over ontogeny in Horokashubuto stream, Hokkaido, northern Japan

AbstractThe relationships between body size and density of salmonids in natural streams have been well studied, and density‐dependent growth (somatic growth) is interpreted as the principal mechanism responsible for the relationship. Moreover, the body size–density relationship is known to vary over ontogeny. However, the relationship has been studied mostly by experimental procedures (e.g. stocking fish into streams), and knowledge about the relationship in natural populations (e.g. no human‐induced input of fish) is still inadequate. In this study, we compared the relationships between body size and density of young‐of‐the‐year (YOY) masu salmon, Oncorhynchus masou, between different ontogenetic stages, that is, fry (spring) and juveniles (autumn). We used the monitoring data of YOY body sizes and densities across three study sites collected from Horokashubuto stream in Hokkaido, northern Japan, from 2014 to 2022. In the juvenile stage, YOY body size correlated negatively with increasing density. This pattern is typical of the density dependence of salmonid growth demonstrated in many previous studies. In the fry stage, however, YOY body size correlated positively with increasing density. This study suggested that the patterns of density dependence vary as functions of the ontogenetic stages of salmonids.

Vertical eDNA distribution of cold-water fishes in response to environmental variables in stratified lake.

In summer, the survival zones of cold-water species are predicted to narrow by both increasing water temperatures from the surface and by expanding hypoxic zones from the lake bottom. To examine how the abundance of cold-water fishes changes along environmental gradients, we assessed the vertical environmental DNA (eDNA) distributions of three salmonid species which may have different water temperature tolerances during both stratification and turnover periods using quantitative PCR (qPCR). In addition, we examined on the vertical distribution of diverse fish fauna using an eDNA metabarcoding assay. The results suggested that the kokanee salmon (Oncorhynchus nerka) eDNA were abundant in deep, cold waters. On the other hand, rainbow trout (O. mykiss) eDNA were distributed uniformly throughout the water column, suggesting that they may have high water-temperature tolerance compared with kokanee salmon. The eDNA concentrations of masu salmon (O. masou) were below the detection limit (i.e., <10 copies μL-1) at all stations and depths and hence could not be quantified during stratification. Together with the finding that the eDNA distributions of other prey fish species were also constrained vertically in species-specific ways, our results suggest that climate change will result in substantial changes in the vertical distributions of lake fish species and thus affect their populations and interactions.

The cytokine genes of Oncorhynchus masou formosanus include a defective interleukin-4/13A gene

Oncorhynchus masou formosanus (Formosa landlocked salmon) is a critically endangered salmonid fish endemic to Taiwan. To begin to understand how its drastic change in lifestyle from anadromous to exclusively river-dwelling is reflected in its immune genes, we characterized the genes encoding six cytokines (IL-2A, IL-2B, IL-4/13A, IL-4/13B1, IL-4/13B2, and IL-17A/F2a) important for T cell responses as no genomic data is available for this fish. Interestingly, all genes appeared homozygous indicative of a genetic bottleneck. The IL2 and IL17A/F2a genes and their products are highly similar to their characterized homologs in Oncorhynchus mykiss (rainbow trout) and other salmonid fish. Two notable differences were observed in IL4/13 family important for type 2 immune responses. First, O. m. formosanus carries not only one but two genes encoding IL-4/13B1 proteins and expansions of these genes are present in other salmonid fish. Second, the OmfoIL4/13A gene carries a 228 bp deletion that results in a premature stop codon and hence a non-functional IL-4/13A cytokine. This suggests a reduced ability for T cell responses against parasitic infections in this species.

Diet Diversity of the Fluviatile Masu Salmon, Oncorhynchus masou (Brevoort 1856) Revealed via Gastrointestinal Environmental DNA Metabarcoding and Morphological Identification of Contents.

Masu salmon, Oncorhynchus masou (Brevoort 1856), a commercially important fish species endemic to the North Pacific Ocean, attained national second-level protected animal status in China in 2021. Despite this recognition, knowledge about the trophic ecology of this fish remains limited. This study investigated the diet diversity of fluviatile Masu salmon in the Mijiang River, China, utilizing the gastrointestinal tract environmental DNA (GITeDNA) metabarcoding and morphological identification. The results revealed a diverse prey composition, ranging from terrestrial and aquatic invertebrates to small fishes. The fluviatile Masu salmon in general consumed noteworthily more aquatic prey than terrestrial prey. There were much more prey taxa and a higher diet diversity detected by GITeDNA metabarcoding than by morphological identification. GITeDNA metabarcoding showed that larger and older Masu salmon consumed significantly more terrestrial insects than aquatic prey species did, with 7366 verses 5012 sequences in the group of ≥20 cm, 9098 verses 4743 sequences in the group of ≥100 g and 11,540 verses 729 sequences in the group of age 3+. GITeDNA metabarcoding also showed size- and age-related diet diversity, indicating that the dietary niche breadth and trophic diversity of larger and older Masu salmon increased with food resources expanding to more terrestrial prey. Terrestrial invertebrates of riparian habitats play a vital role in the diet of fluviatile Masu salmon, especially larger individuals, highlighting their importance in connecting aquatic and terrestrial food webs. Conservation plans should prioritize the protection and restoration of riparian habitats. This study advocates the combined use of GITeDNA metabarcoding and morphological observation for a comprehensive understanding of fish diet diversity.

Resource partitioning between non-native white-spotted charr and native red-spotted masu salmon in Shikoku, southwestern Japan: population- and individual-level analyses

Resource partitioning between non-native white-spotted charr and native red-spotted masu salmon in Shikoku, southwestern Japan: population- and individual-level analyses

Impacts of Temperature on the Growth, Feed Utilization, Stress, and Hemato-Immune Responses of Cherry Salmon (Oncorhynchus masou).

Cherry salmon (Oncorhynchus masou) hold commercial value in aquaculture, and there is a need for controlled laboratory studies to isolate the specific effects of temperature on their growth, feeding, and well-being. We examined the effects of different temperatures (10 °C, 14 °C, 18 °C, and 22 °C) on juvenile cherry salmon (average mass 29.1 g) in triplicate tanks per treatment over eight weeks. The key parameters assessed included growth rate, feed efficiency, stress response, and hemato-immune responses. Our objectives were to determine the most and less favorable temperatures among the four designated temperatures and to assess the adverse effects associated with these less favorable temperatures. The results showed that body weight, growth rates, feed intake, and feed efficiency were significantly higher at 10 °C and 14 °C compared to 18 °C and 22 °C. Reduced appetite and feeding response were observed at 22 °C. Red blood cell parameters were significantly lower at 22 °C. At 10 °C, the results showed significantly increased plasma cortisol levels, gill Na+/K+-ATPase activity, body silvering, and decreased condition factors, suggesting potential smoltification. The potential smoltification decreased with increasing temperatures and disappeared at 22 °C. Furthermore, the plasma lysozyme concentrations significantly increased at 18 °C and 22 °C. In conclusion, our study identifies 10 °C and 14 °C as the temperatures most conducive to growth and feed performance in juvenile cherry salmon under these experimental conditions. However, temperatures of 22 °C or higher should be avoided to prevent compromised feeding, reduced health, disturbed immune responses, impaired growth, and feed performance.

Infection status of masu salmon Oncorhynchus masou by the parasite Salmonema cf. ephemeridarum (Nematoda, Cystidicolidae).

Nematodes that parasitize salmonids are found in both seawater and freshwater. Unlike seawater species such as those in family Anisakidae, freshwater species have not been well studied. In particular, the influences of these nematodes on the body condition of salmonids remain unclear. We studied the effects of Salmonema cf. ephemeridarum on the body condition of masu salmon Oncorhynchus masou. We found a positive relationship between the number of parasites and fish fork length. In contrast, we found a negative relationship between the body condition (condition factor) of fish and the number of parasites. These results suggest that nematode infection could affect host energy reserves for future growth.