Dispersal of wild and domestic masu salmon fry (Oncorhynchus masou) in an artificial channel

The day and night pattern of upstream and downstream dispersal of masu salmon fry of wild and domestic origin was compared in artificial channels (45 m long), for two ages of planting: unfed alevins and eyed eggs. Early dispersal was important for the wild stock (48–50%) compared with the domestic one (16–36%). More wild fry moved downstream than upstream, and more domestic fry dispersed upstream. Upstream movement in wild and domestic fry was more active by day than by night, except for wild fry planted as eyed eggs, where upstream migration was higher at night. In contrast, downstream movement in wild and domestic fry was more common by night than by day, but daylight catches were not negligible for the wild stock.     

Genetic structure of masu salmon (Oncorhynchus masou) populations in Hokkaido, northernmost Japan, inferred from mitochondrial DNA variation

Genetic structure of masu salmon Oncorhynchus masou populations in Hokkaido was examined by analysing mtDNA NADH dehydrogenase subunit 5 gene (561 bp) of 382 individuals collected from 12 rivers, in which there were no records of artificial release. Analysis of molecular variance showed that between groups level and between populations within-group level explained each c. 10% of genetic variance. In neighbour-joining tree, four populations located in southern Hokkaido were clustered into a single group; however, other populations did not form any clear clusters. Fu's F _S, Tajima's D and a mismatch distribution test indicated a sudden expansion of population in the entire population of Hokkaido and the northernmost population of Chiraibetsu, which was genetically close to the southern Hokkaido group. The Sea of Japan and southern rivers, including those of southern Hokkaido, seem to have served as refugia for masu salmon during glacial periods, and their dispersal and straying in interglacial periods affected the genetic structure of masu salmon populations in Hokkaido.     

Effects of melatonin feeding on smoltification in masu salmon (Oncorhynchus masou)

Influences of photoperiod on plasma melatonin profiles and effects of melatonin administration on long-day-induced smoltification in masu salmon (Oncorhynchus masou) were investigated in order to reveal the roles of melatonin in the regulation of smoltification in salmonids. Under light-dark (LD) cycles, plasma melatonin levels exhibited daily variation, with higher values during the dark phase than during the light phase. The duration of nocturnal elevation under short photoperiod (LD 8:16) was longer than that under long photoperiod (LD 16:8). Melatonin feeding (0.01, 0.1 and 1 mg/kg body weight) elevated plasma levels of melatonin in a dose-dependent manner for at least 7 h but not for 24 h. When masu salmon reared under short photoperiod were exposed to long photoperiod (LD 16:8) and fed melatonin (1 mg/kg body weight) 7 hours before the onset of darkness, a significantly smaller proportion of smolts appeared in the melatonin-fed group after 32 days than in the control group. However, after 59 days of the treatment, there was no difference in the proportion of smolts between the control and melatonin-treated groups. Thus, melatonin feeding mimicked the effects of short photoperiod, which delays but does not completely suppress smoltification. These results indicate that the day length is transduced into changes in the duration of nocturnal elevation in plasma melatonin levels, and that artificial modification of the plasma melatonin pattern possibly delays the physiological processes of smoltification induced by long-day photoperiodic treatment.     

Proteomic analysis of inviable salmonid hybrids between female masu salmon Oncorhynchus masou masou and male rainbow trout Oncorhynchus mykiss during early embryogenesis

Early embryos of inviable hybrids between female masu salmon Oncorhynchus masou masou and male rainbow trout Oncorhynchus mykiss at 9, 12, 15 and 20 days after fertilization were examined for protein expression profiles. A total of 44 proteins, mostly down-regulated products of house-keeping genes and those involved in nucleic acid metabolism or chromatin replication, were identified in hybrid embryos by mass spectrometry analysis and protein database searching. The identified down-regulated proteins may be responsible for the inviability in the hybrids.     

Henneguya cartilaginis n. sp. (Myxozoa: Myxosporea) in the head cartilage of masu salmon Oncorhynchus masou masou

Henneguya cartilaginis n. sp. (Myxozoa: Myxosporea) is described from wild masu salmon Oncorhynchus masou masou in Hokkaido, Japan. H. cartilaginis n. sp. produced white cysts, up to 3mm in size, in the head of masu salmon. Infected fish exhibited cranial protrusion due to the cysts. Spores (11.4 × 8.6μm) of H. cartilaginis n. sp. were egg-shaped with the posterior end more pointed and possessed two caudal appendages (34.2μm average length). Histological observations revealed that large plasmodia possessing fine fibrous pseudopodia on the surface developed in the head cartilage. H. cartilaginis n. sp. resembles H. cerebralis, which was described from the cranial cartilage of Kosogol grayling Thymallus nigrescens in Mongolia. However, they were distinguishable by spore morphology. Molecular analysis of the 18S rDNA sequences indicated that H. cartilaginis n. sp. was most closely related to Henneguya zschokkei, H. nuesslini and H. salminicola of salmonid fish, with genetic similarities of 95.3%, 95.1% and 93.9%, respectively. Based on these differences in spore morphology, molecular data, the site of infection and geographical distribution, the present species is considered to be a new species.     

A visceral mycosis in cultured masu salmon (Oncorhynchus masou) caused by a species of Ochroconis

Ochronconis sp. infection was found in masu salmon (Oncorhynchus masou) with visceral mycosis in Japan. The external and internal clinical signs were reddening of the anal area, swelling of the abdomen due to accumulation of ascitic fluid in the abdominal cavity and extensive swelling of the posterior kidney. Many pale brown, septate hyphae were found in the kidney by direct microscopical examination; these were usually not found in the other organs of infected fish. Histopathological examination of the kidney revealed large granulomas with the fungal hyphae and giant cells. The isolated fungus was identified as a species of the genus Ochroconis and was compared with O. tshawytschae, a known fish pathogen. Based on morphological and growth characteristics, we believe that these cases resulted from infection with a different species.     

Purification of serum precursor proteins to vitelline envelope (choriogenins) in masu salmon,Oncorhynchus masou

Three vitelline envelope-related proteins (VERPs), very-high-molecular-weight VERP (vhVERP), high-molecular-weight VERP (hVERP) and low-molecular-weight VERP (lVERP) were purified from female masu salmon serum. The apparent molecular weights of vhVERP, hVERP and lVERP, in their native state, were 520, 88 and 54 kDa, respectively, by gel-filtration chromatography. Very-high-molecular-weight VERP comprises two subunits, corresponding to 175 and 126 kDa. On SDS-PAGE, hVERP and lVERP migrate at 53 and 47 kDa, respectively. Amino acid analysis of vhVERP and hVERP showed that they share a high content of glutamic acid and proline. By contrast, lVERP is rich in glutamic acid and asparatic acid. These features are in good agreement with the amino acid composition of the vitelline envelope. Immuno-biochemical analysis suggested that vhVERP is derived from hVERP by polymerization and/or aggregation. Antibodies against hVERP and lVERP specifically immunostained the vitelline envelope and liver of female masu salmon. In addition, both hVERP and lVERP were induced in the serum of estrogen-treated male fish. Taken together, it is suggested that hVERP and lVERP are homologous molecules with choriogenin H and choriogenin L in medaka, respectively. These results indicate that hVERP and lVERP are precursor proteins to the vitelline envelope (choriogenins) in masu salmon.     

Latitudinal variation in sexual size dimorphism of sea-run masu salmon, Oncorhynchus masou

Sexual size dimorphism (SSD), a difference in body size between the sexes, occurs in many animal species. Although the larger sex is often considered invariable within species, patterns of selection may result in interpopulation variation or even reversal of SSD. We evaluated correlations between latitude and female body size, male body size, and relative body size (male body size/female body size) in 22 populations (ranging from 37 degrees N to 49 degrees N) of sea-run masu salmon (Oncorhynchus masou) that spawn in rivers along the Sea of Japan coast. Male size and the relative body size increased with latitude, but female size did not correlate with latitude. In addition, increase in male size with latitude was sufficient to result in a reversal of SSD, the switch-point being around 45 degrees N. We suggest that the positive correlation between latitude and male size is due to increasing operational sex ratios or sexual selection on sea-run male body size that result from sex-biased patterns of anadromy. In conclusion, our study provides the first example of predictable geographic variation in SSD shaped by apparent patterns of sexual selection.     

Functional analysis of thermo-sensitive TRPV1 in an aquatic vertebrate,masu salmon (Oncorhynchus masou ishikawae)

Transient receptor potential vanilloid 1 (TRPV1) is mainly expressed in nociceptive primary sensory neurons and acts as a sensor for heat and capsaicin. The functional properties of TRPV1 have been reported to vary among species and, in some cases, the species difference in its thermal sensitivity is likely to be associated with thermal habitat conditions. To clarify the functional properties and physiological roles of TRPV1 in aquatic vertebrates, we examined the temperature and chemical sensitivities of TRPV1 in masu salmon (Oncorhynchus masou ishikawae, Om) belonging to a family of salmonids that generally prefer cool environments. First, behavioral experiments were conducted using a video tracking system. Application of capsaicin, a TRPV1 agonist, induced locomotor activities in juvenile Om. Increasing the ambient temperature also elicited locomotor activity potentiated by capsaicin. RT-PCR revealed TRPV1 expression in gills as well as spinal cord. Next, electrophysiological analyses of OmTRPV1 were performed using a two-electrode voltage-clamp technique with a Xenopus oocyte expression system. Heat stimulation evoked an inward current in heterologously expressed OmTRPV1. In addition, capsaicin produced current responses in OmTRPV1-expressing oocytes, but higher concentrations were needed for its activation compared to the mammalian orthologues. These results indicate that Om senses environmental stimuli (heat and capsaicin) through the activation of TRPV1, and this channel may play important roles in avoiding environments disadvantageous for survival in aquatic vertebrates.     

Molecular cloning and characterization of three distinct choriogenins in masu salmon, Oncorhynchus masou

Three cDNAs, each encoding a different choriogenin (Chg), were isolated from a female masu salmon (Oncorhynchus masou) liver cDNA library. Two of the cDNA clones, Chg Halpha and Chg Hbeta, showed a close relationship and contained the typical domains of zona pellucida (ZP) B genes in fish, namely proline and glutamine rich repeats, a trefoil factor family domain, and a ZP domain. Specific antibodies against recombinant Chg H products (rmHalpha and rmHbeta) were generated to elucidate the relationship between the Chg H cDNAs and two types of serum Chg H protein, which were previously purified and characterized, and designated as very-high-molecular-weight vitelline envelope-related protein (vhVERP) and Chg H of masu salmon. The immunobiochemical analyses revealed that the Chg Halpha and Chg Hbeta clones encoded vhVERP and Chg H proteins, respectively. The third cDNA clone (Chg L) appeared to be a ZPC gene and, by mapping the N-terminal sequence of purified Chg L, was shown to encode serum Chg L protein. Various types of heteromultimer of the three Chgs were identified immunologically as high molecular weight chorion components, indicating the involvement of complex heterodimerization of multiple Chgs in the construction of chorion architecture in masu salmon.     

Estimation of migratory history in female masu salmon Oncorhynchus masou (Brevoort, 1856) with the growth hormone pseudogene

To estimate the migration timing, length, and location of females with the growth hormone pseudogene (GHp), trace element analyses were performed on the otoliths of 40 wild adult masu salmon, Oncorhynchus masou (12 males, 19 normal females, and nine females with GHp [all fish were 3‐ or 4‐years‐old]) caught during the reproductive season (September) from 2004 to 2010 in the Atsuta River in Japan. Sr concentrations at the end of the otolith translucent area formed during the winter suddenly increased in all individuals, suggesting that they had migrated in the spring. In addition, the estimated migration length and location of females with GHp indicated that they had spent a fixed period of 1 year in the ocean, suggesting that females with GHp have a similar migratory history as that of normal females.     

Ascending speed and nocturnal activity of adult masu salmon (Oncorhynchus masou Brevoort) during upstream migration

We released five adult masu salmon (Oncorhynchus masou) tagged with external transmitters to track their ascending behaviour. The signals of all specimens were recorded in the upper area of the river system. Two patterns of ascending behaviour were recognized: ascending upward immediately after release and ascending during increased river discharge. The fastest ascending speed was about 1000 m h^–1. Active movements were detected at night. The signal recording duration at each receiver for each fish was generally brief. Most fish did not stay at the pools where the receivers were installed.     

Genetic diversity and differentiation of masu salmon (Oncorhynchus masou masou) between and within cultured populations inferred from microsatellite DNA analysis

Masu salmon, Oncorhynchus masou masou, is one of the most valuable fishery species that has been introduced to China, though to date no studies on the genetic diversity and genetic relationship among hatchery populations has been performed with molecular markers. We undertook such a study and sampled 120 individuals from three hatchery stocks and analyzed 20 microsatellite loci. All loci were polymorphic and a total of 91 alleles were detected. A relatively low level of genetic diversity was revealed with effective number of allele of 3.1094, 3.3299 and 3.1894 and expected heterozygosity of 0.6600, 0.6648 and 0.6638 in the three stocks, respectively. Deviations from Hardy-Weinberg equilibrium were found due to heterozygote deficit. Accordingly, evidence of genetic bottlenecks were found in the three stocks. An individual assignment test demonstrated that 85% of individuals were correctly assigned into their original stocks. Pairwise Fst revealed that significant differentiation occurred between these three stocks. The results of the study indicated that disequilibrium of genetic structure and differentiation has occurred in all three stocks. This information collectively provides a basis for measures to avoid of loss of genetic diversity and introgression in Chinese aquaculture.     

Phylogenetic position and genetic diversity of masu salmon Oncorhynchus masou in the Mijiang River,China

Ichthyological Research -     

Genetic diversity and differentiation of masu salmon (Oncorhynchus masou masou) between and within cultured populations inferred from microsatellite DNA analysis

Masu salmon,Oncorhynchus masou masou,is one of the most valuable fishery species that has been introduced to China,though to date no studies on the genetic diversity and genetic relationship among hatchery populations has been performed with molecular markers.We undertook such a study and sampled 120 individuals from three hatchery stocks and analyzed 20 microsatellite loci.All loci were polymorphic and a total of 91 alleles were detected.A relatively low level of genetic diversity was revealed with effective number of allele of 3.1094,3.3299 and 3.1894 and expected heterozygosity of 0.6600,0.6648 and 0.6638 in the three stocks,respectively.Deviations from Hardy-Weinberg equilibrium were found due to heterozygote deficit.Accordingly,evidence of genetic bottlenecks were found in the three stocks.An individual assignment test demonstrated that 85% of individuals were correctly assigned into their original stocks.Pairwise Fst revealed that significant differentiation occurred between these three stocks.The results of the study indicated that disequilibrium of genetic structure and differentiation has occurred in all three stocks.This information collectively provides a basis for measures to avoid of loss of genetic diversity and introgression in Chinese aquaculture.     

Effects of woody debris on the habitat of juvenile masu salmon (Oncorhynchus masou) in northern Japanese streams

1. The effects of woody debris on stream habitat of juvenile masu salmon ( Oncorhynchus masou ) were examined at two spatial scales, stream reach and channel unit, for first to thirdorder tributaries of the Teshio River in northern Hokkaido, Japan. The fortyeight study reaches were classified into three distinct types: coarsesubstrate steppool (CSP), coarsesubstrate poolriffle (CPR) and finesubstrate poolriffle (FPR) reaches. Each reach type included reaches with different riparian settings, broadly classified as forest (relatively undisturbed forest and secondary forest after fires) or grassland (bamboo bushland and pasture).
2. The reachscale analyses showed that neither total pool volume nor pooltopool spacing was correlated with woody debris abundance in any of the three reach types. Masu salmon density was positively correlated with both woodydebris cover area and total cover area, but not with total pool volume in the reaches.
3. Channelunitscale analyses revealed that woody debris reduced nonpool velocity, increased pool depth and retained fine sediment in pools in FPR reaches, where the size of woody debris was very large relative to the substrate material size. However, woody debris did not influence any of the hydraulic variables (depth, velocity, substrate) in either nonpools or pools of CSP and CPR reaches. Habitat use by masu salmon in nonpools or pools was affected by woodydebris cover area or total cover area rather than by hydraulic variables in any of the reach types.
4. The effects of woody debris on habitat at the reach and channelunit scales in the study area were less than those indicated by previous work in the Pacific Northwest, North America, owing to the relatively small size of the riparian trees. However, the overall results suggested that woody debris in the study area contributed to masu salmon habitat by providing cover at the smaller, microhabitat scale.     

New polymorphic microsatellite markers for the masu salmon (Oncorhynchus masou masou) from Korea and their application to wild and hatchery populations

Masu salmon, Oncorhynchus masou masou, is an economically important fish species in the Far East and occurs in two life history forms: sea-run migratory (anadromous) and freshwater resident (non-anadromous). The non-anadromous form has recently become a popular freshwater food and game fish during a well-known Korean winter festival. However, the genetic background of this species remains largely unknown, partly due to a lack of molecular genetic markers. In this study, we developed new polymorphic microsatellite markers for masu salmon using next-generation sequencing technology. From 40 primer sets, 11 primer sets (27.5% of the primer sets selected) were successfully amplified with 106 alleles (range 2–9) in 64 individuals from different populations: two wild and one hatchery. Observed and expected heterozygosities ranged from 0.304 to 0.947 and 0.278 to 0.865, respectively. Significant departures from the Hardy–Weinberg equilibrium were detected for four markers (OMM11, OMM17, OMM28, and OMM33) in a single population. All pair-wise F_ST values were highly significant between the wild and hatchery populations (range 0.084–0.183, P < 0.0001). We identified a set of robust microsatellite markers that worked well even in formalin-fixed samples, which will be suitable for biogeographical and population structure analyses of the masu salmon.     

Riverine, estuarine and marine migratory behaviour and physiology of wild and hatchery-reared coho salmon Oncorhynchus kisutch (Walbaum) smolts descending the Campbell River, BC, Canada

Eighty coho salmon Oncorhynchus kisutch smolts (40 wild and 40 hatchery-reared) were surgically implanted with acoustic transmitters and released into the Quinsam River over 2 days. Differences in physiology, travel time and migratory behaviour were examined between wild and hatchery-reared fish. In addition, tagged and control fish of both wild and hatchery-reared stock were raised for 3 months following surgery to compare survival and tag retention. Detection ranges of the acoustic receivers were tested in the river, estuary and ocean in a variety of flow conditions and tide levels. Receivers were placed in the river, estuary and up to 50 km north and south from the river mouth in the marine environment. Wild smolts were significantly smaller by mass, fork length and condition factor than hatchery-reared smolts and exhibited significantly higher levels of sodium, potassium and chloride in their blood plasma than hatchery-reared smolts. The gill Na⁺K⁺-ATPase activity was also significantly higher in the wild coho smolts at the time of release. Ninety-eight per cent of wild and 80% of hatchery-reared fish survived to the estuary, 8 km downstream of the release site. No difference was found in migration speed, timing or survival between smolts released during daylight and those released after dark. Wild smolts, however, spent less time in the river and estuary, and as a result entered the ocean earlier than hatchery-reared smolts. Average marine swimming speeds for wild smolts were double those of their hatchery-reared counterparts. While hatchery smolts dispersed in both a northward and southward direction upon entering the marine environment, the majority of wild smolts travelled north from the Campbell River estuary. The wild coho salmon smolts were more physiologically fit and ready to enter sea water than the hatchery-reared smolts, and as a result had higher early survival rates and swimming speeds.     

Spatial distribution of juvenile masu salmon (Oncorhynchus masou) with incongruent genotypic and phenotypic sex in Hokkaido, Japan

To clarify the spatial distribution of masu salmon Oncorhynchus masou with incongruent genotypic and phenotypic sex, we compared the phenotypic sex determined by gonad analysis with the genotypic sex identified by a male-specific genetic marker, Growth hormone pseudogene (GHp), in 584 individuals inhabiting 23 rivers in Hokkaido, Japan. We found that the genotypic sex is different from the phenotypic sex for 39 individuals in 10 rivers. Among them, 35 individuals (89.7?%) were GHp?+?females phenotypically. The opposite feature was noted in the remaining four individuals (10.3?%). This study also revealed that GHp?+?phenotypic females are distributed mainly in the Sea of Japan, whereas GHp- phenotypic male tend to appear in the Sea of Okhotsk. In addition, there was a significant negative correlation between the distance from the Bishabetsu River, where frequency of GHp?+?phenotypic females was the highest, and the frequency of those individuals in each river. These results suggest that the distribution of masu salmon with incongruent genotypic and phenotypic sex would be related to the movement of individuals among the populations and regions.