Seawater Adaptation Research Articles

Genomic insights into the seawater adaptation in Cyprinidae

BackgroundCyprinidae, the largest fish family, encompasses approximately 367 genera and 3006 species. While they exhibit remarkable adaptability to diverse aquatic environments, it is exceptionally rare to find them in seawater, with the Far Eastern daces being of few exceptions. Therefore, the Far Eastern daces serve as a valuable model for studying the genetic mechanisms underlying seawater adaptation in Cyprinidae.ResultsHere, we sequenced the chromosome-level genomes of two Far Eastern daces (Pseudaspius brandtii and P. hakonensis), the two known cyprinid fishes found in seawater, and performed comparative genomic analyses to investigate their genetic mechanism of seawater adaptation. Demographic history reconstruction of the two species reveals that their population dynamics are correlated with the glacial-interglacial cycles and sea level changes. Genomic analyses identified Pseudaspius-specific genetic innovations related to seawater adaptation, including positively selected genes, rapidly evolving genes, and conserved non-coding elements (CNEs). Functional assays of Pseudaspius-specific variants of the prolactin (prl) gene showed enhanced cell adaptation to greater osmolarity. Functional assays of Pseudaspius specific CNEs near atg7 and usp45 genes suggest that they exhibit higher promoter activity and significantly induced at high osmolarity.ConclusionsOur results reveal the genome-wide evidence for the evolutionary adaptation of cyprinid fishes to seawater, offering valuable insights into the molecular mechanisms supporting the survival of migratory fish in marine environments. These findings are significant as they contribute to our understanding of how cyprinid fishes navigate and thrive in diverse aquatic habitats, providing useful implications for the conservation and management of marine ecosystems.

Transcriptional regulation of esophageal, intestinal, and branchial solute transporters by salinity, growth hormone, and cortisol in Atlantic salmon.

In marine habitats, Atlantic salmon (Salmo salar) imbibe seawater (SW) to replace body water that is passively lost to the ambient environment. By desalinating consumed SW, the esophagus enables solute-linked water absorption across the intestinal epithelium. The processes underlying esophageal desalination in salmon and their hormonal regulation during smoltification and following SW exposure are unresolved. To address this, we considered whether two Na+ /H+ exchangers (Nhe2 and -3) expressed in the esophagus contribute to the uptake of Na+ from lumenal SW. There were no seasonal changes in esophageal nhe2 or -3 expression during smoltification; however, nhe3 increased following 48 h of SW exposure in May. Esophageal nhe2, -3, and growth hormone receptor b1 were elevated in smolts acclimated to SW for 2.5 weeks. Treatment with cortisol stimulated branchial Na+ /K+ -ATPase (Nka) activity, and Na+ /K+ /2Cl- cotransporter 1 (nkcc1), cystic fibrosis transmembrane regulator 1 (cftr1), and nka-α1b expression. Esophageal nhe2, but not nhe3 expression, was stimulated by cortisol. In anterior intestine, cortisol stimulated nkcc2, cftr2, and nka-α1b. Our findings indicate that salinity stimulates esophageal nhe2 and -3, and that cortisol coordinates the expression of esophageal, intestinal, and branchial solute transporters to support the SW adaptability of Atlantic salmon.

Successful delivery of viviparous lantern shark from an artificial uterus and the self-production of lantern shark luciferin.

Our recent success in the long-term maintenance of lantern shark embryos in artificial uterine systems has provided a novel option for the medical treatment of premature embryos for captive viviparous elasmobranchs. The remaining issue with this system is that the embryos cannot survive the abrupt change in the chemical environment from artificial uterine fluid (AUF) to seawater during delivery. To overcome this issue, the present study developed a new protocol for seawater adaptation, which is characterized by a long-term and stepwise shift from AUF to seawater prior to delivery. This protocol was employed successfully, and the specimen survived for more than seven months after delivery, the longest captive record of the species. During the experiment, we unexpectedly detected bioluminescence of the embryonic lantern shark in the artificial uterus. This observation indicates that lantern sharks can produce luciferin, a substance for bioluminescence. This contradicts the recent hypothesis that lantern sharks lack the ability to produce luciferin and use luciferin obtained from food sources.

Exposure to silver impairs the osmoregulatory capability of euryhaline medaka (Oryzias latipes) subjected to salinity changes

The widespread use of silver in nanomaterials has led to increases in environmental contamination, which poses a threat to aquatic animals. Euryhaline fish, which live in environments with fluctuating salinity levels, have strong osmotic regulatory abilities to cope with such changes. This study attempted to investigate how silver affects the osmoregulatory capabilities of euryhaline fish, using medaka (Oryzias latipes) embryos as a model. The embryos were exposed to AgNO3 for 7 d in either fresh water (FW) or seawater (SW), and their mortality, heart rate, morphology, and ionocytes were examined. Results showed that the toxicity of AgNO3 was higher in FW than in SW (50% lethal concentrations (LC50) were 0.17 vs. 1.01 ppm). Although AgNO3 (0.05 and 0.1 ppm) did not significantly change the morphology of embryos, it impaired ionocytes and elevated heart rates in FW. While, AgNO3 (0.1 and 0.5 ppm) did not affect the morphology, ionocytes, or heart rate in SW, it impaired the hypo-osmoregulatory capability and elevated the mortality of embryos that were transferred from FW to SW. At 12 h after SW transfer, ionocytes were severely impaired, and water-drinking behavior was suppressed, resulting in body dehydration and sodium overload. In contrast, AgNO3 did not elevate the mortality of embryos that were transferred from SW to FW. To sum up, the presence of silver in FW during the developmental stage of euryhaline fish could potentially endanger their survival during SW adaptation.

Study of Mechanical and Thermal Properties for Epoxy Grouts Subjected to Seawater Conditioning at Elevated Temperature: Tensile Test and Compressive Test

Pipelines and risers are major transportation of oil and gas. These components are often exposed to extreme marine environmental conditions that can cause pipelines to fail due to corrosion. Hence, epoxy grouts are used as a coating material on the surface of pipelines and risers for supporting the pipe structure and corrosion prevention. In this case, the efficiency of epoxy grout is important in pipeline rehabilitation. Research on the adaptation of seawater, temperature, and epoxy grouts is carried out to determine the changes in mechanical and thermal properties of the material. The preparation of epoxy grouts was prepared with a mixture of epoxy resins, epoxy hardeners, and aggregates. After the sample compounds have been flattened, the mixture will be put into the mold. The seawater aging process was performed for seven days before conducting laboratory experiments. After the seawater aging process, there were physical changes on the surface of the epoxy grouts which are the epoxy grout become harder and there were voids that can act as stress concentration points that may cause micro-crack on the specimen. Next, tensile (ASTM 638) experiments were performed at different temperatures of 27°C, 40°C, 52°C, 72°C, 80°C and 100°C while the result of compression experiment is based on the literature critique of past journals. Based on the result, the young modulus and ultimate tensile strength of the epoxy grouts are decreased with increasing temperature. The increment of temperature causes the epoxy grouts to become weaker due to the changes in the polymer matrix structure of the epoxy grouts which lead to the failure at low load and shortens the life of the material. In addition, epoxy grouts are amorphous materials where the glass transition temperature determine the mechanical and physical properties of epoxy grouts. Therefore, the results of the study found that the epoxy grouts changed its mechanical properties from brittle to ductile when the temperature is at 72°C. From the comparison between samples immersed and not immersed in seawater, there is a decrease in Young’s modulus, ultimate tensile stress and strain due to the effect of seawater reaction. In conclusion, the increment in temperature and seawater adaptation factors affect the strength and durability of epoxy grouts.

Interaction of temperature and feed ration on male postsmolt maturation of Atlantic salmon (Salmo salar L.)

Early maturation of Atlantic salmon male postsmolts is undesirable in aquaculture due to its negative impact on growth, welfare and seawater adaptation, however it is an increasing problem under intensive rearing conditions. The effect of temperature and feeding ration on sexual maturation during the induction of smoltification was tested with a 3 × 2 factorial design including three temperatures (8, 12.5 and 18 °C) and two feeding rations (100% and 67%), from 28 October 2018 to 30 May 2019. Fish in the six resulting treatments (1800 parr, initial mean weight 23.1 ± 7.2 g) were reared in flow-through under continuous light (LD24:0) except for a 5-week winter signal (LD12:12) introduced on 4 February 2019 to induce smoltification. A major effect of temperature determining the life history pattern of each group was observed. At 18 °C, a majority of the males were maturing in May under both rations (100% males in 18–100% and 88.9% males in 18–67%). Fish at 18 °C experienced an early activation of the BPG (Brain-Pituitary-Gonad) axis and displayed very poor signs of smoltification. At 12.5 °C, early maturation was also present, although at lower proportion than at 18 °C and more dependent upon feed ration (40% of males maturing in 12.5–100%, 8.6% in 12.5–67%). Salmon at 12.5 °C displayed the best growth within each feed regime, some early signs of smolting but subsequent low Na+, K+- ATPase activity after the winter signal. At 8 °C, salmon showed best signs of smoltification and lowest growth, while no males matured. The feed restriction had a minor effect on maturation at 18 °C but a greater impact at 12.5 °C, evidenced by significant differences between 12.5–100% and 12.5–67% in gonadosomatic index, plasma 11-ketotestosterone, insulin-like growth factor-I, and pituitary fshb (follicle stimulating hormone β-subunit gene) transcription. Onset of maturation was associated with the photoperiod change from short to long daylength, which probably also induced a slight activation of the BPG axis in non-maturing groups. Our results suggest that postsmolt producers must be cautious when using sustained high water temperatures, or the risk of early maturation and poor hypo-osmoregulatory performance will increase significantly. In addition, reducing the feeding ration will not help decrease incidence of maturation without significantly affecting growth.

Differential Expression of miRNAs and Their Predicted Target Genes Indicates That Gene Expression in Atlantic Salmon Gill Is Post-Transcriptionally Regulated by miRNAs in the Parr-Smolt Transformation and Adaptation to Sea Water.

Smoltification (parr-smolt transformation) is a complex developmental process consisting of developmental changes that lead to remodeling of the Atlantic salmon gill. Here, the expression changes of miRNAs and mRNAs were studied by small-RNA sequencing and microarray analysis, respectively, to identify miRNAs and their predicted targets associated with smoltification and subsequent sea water adaptation (SWA). In total, 18 guide miRNAs were identified as differentially expressed (gDE miRNAs). Hierarchical clustering analysis of expression changes divided these into one cluster of 13 gDE miRNAs with decreasing expression during smoltification and SWA that included the miRNA-146, miRNA-30 and miRNA-7132 families. Another smaller cluster that showed increasing expression consisted of miR-101a-3p, miR-193b-5p, miR-499a-5p, miR-727a-3p and miR-8159-5p. The gDE miRNAs were predicted to target 747 of the genes (DE mRNAs), showing expression changes in the microarray analysis. The predicted targets included genes encoding NKA-subunits, aquaporin-subunits, cystic fibrosis transmembrane conductance regulator and the solute carrier family. Furthermore, the predicted target genes were enriched in biological processes associated with smoltification and SWA (e.g., immune system, reactive oxygen species, stress response and extracellular matrix organization). Collectively, the results indicate that remodeling of the gill involves the post-transcriptional regulation of gene expression by the characterized gDE miRNAs.

Expression Analysis in Atlantic Salmon Liver Reveals miRNAs Associated with Smoltification and Seawater Adaptation.

Simple SummarySmoltification is a developmental process that preadapts Atlantic salmon for a life in seawater. Suboptimal smoltification and poor timing of transfer to seawater is associated with increased mortality. MicroRNAs (miRNAs) are small non-coding genes. They regulate gene expression post-transcriptionally as part of the miRNA induce silencing complex (miRISC) where they guide miRISC to particular mRNAs (target genes). The aim of this study was to identify Atlantic salmon miRNAs expressed in liver that are associated with smoltification and adaptation to seawater as well as to predict their target genes. In total, 62 guide miRNAs were identified, and by their expression patterns they were clustered into three groups. Target gene predictions followed by gene enrichment analysis of the predicted targets indicated that the guide miRNAs were involved in post-transcriptional regulation of important smoltification associated biological processes. Some of these were energy metabolism, protein metabolism and transport, circadian rhythm, stress and immune response. Together, the results indicate that certain miRNAs are involved in the regulation of many of the important changes occurring in the liver during this developmental transition.Optimal smoltification is crucial for normal development, growth, and health of farmed Atlantic salmon in seawater. Here, we characterize miRNA expression in liver to reveal whether miRNAs regulate gene expression during this developmental transition. Expression changes of miRNAs and mRNAs was studied by small-RNA sequencing and microarray analysis, respectively. This revealed 62 differentially expressed guide miRNAs (gDE-miRNAs) that could be divided into three groups with characteristic dynamic expression patterns. Three of miRNA families are known as highly expressed in liver. A rare arm shift was observed during smoltification in the Atlantic salmon-specific novel-ssa-miR-16. The gDE-miRNAs were predicted to target 2804 of the genes revealing expression changes in the microarray analysis. Enrichment analysis revealed that targets were significantly enriched in smoltification-associated biological process groups. These included lipid and cholesterol synthesis, carbohydrate metabolism, protein metabolism and protein transport, immune system genes, circadian rhythm and stress response. The results indicate that gDE-miRNAs may regulate many of the changes associated with this developmental transition in liver. The results pave the way for validation of the predicted target genes and further study of gDE-miRNA and their targets by functional assays.

Significance of sea entry pathway of chum salmon Oncorhynchus keta fry, inferred from the differential expressions of Na+,K+-ATPase α-subunit genes in the gills

Na+,K+-ATPase (NKA) α-subunit 1a (α1a) and 1b (α1b) gene expressions in the gills are changeable in response to ambient salinity in a few salmonids. In this study, the expressions were compared among ambient salinities and used to infer sea entry migration of chum salmon Oncorhynchus keta fry. The expression of α1a decreased from the 2 days after seawater (SW) transfer from freshwater (FW) and was significantly lower in SW-acclimated fry than that in FW-fry. On the other hand, the expression of α1b peaked on the first to second day after SW transfer and then settled to a level 2-fold higher than in FW-fry. In fry caught in the waterfronts of the beaches, the expression levels were quite similar to those on the first and second days after SW transfer, whereas, in fry caught off beach, the expressions were identical to those of SW-acclimated fry. These suggest that fry adapt to SW with moving along the shoal in the bay, and move to off beach after completing SW adaptation. One of the physiological significances in a usage of waterfront may be to transform the gills to SW type. Only fry on the 2 days after SW transfer failed to exhibit condition factor-dependency of burst swimming, probably due to physiological perturbation, which may be related to poor predation avoidance. The physiological approach used in this study inferred sea entry migration of fry; furthermore, it shows the possible significance of adaptation to SW in the shoal is to reduce predation risk.

Five-Month Incubation of Viviparous Deep-Water Shark Embryos in Artificial Uterine Fluid

In this study, we developed an extra-uterine life-support system (artificial uterus) for viviparous elasmobranchs. Using this system, we maintained two embryonic specimens of the slendertail lantern shark (Etmopterus molleri) for approximately 5 months, the longest published record of the captive maintenance of the embryos of this species. This system is characterized by the use of a urea-containing artificial incubation fluid, of which the salinity and osmotic pressure are approximately equivalent to those of shark blood plasma. We hypothesized that this fluid reduces the salinity gradient between embryonic blood plasma and the environmental fluid, which thereby contributes to countering the low osmoregulatory capacity of viviparous shark embryos. However, the incubated specimens subsequently died during the process of seawater adaptation following “artificial birth.” Accordingly, information pertaining to uterine chemical dynamics throughout the gestation period is necessary for the design of a more effective incubation system. This technique will be relevant and may find application in conservation breeding at public aquaria in the future.

Whole-Genome Transcript Expression Profiling Reveals Novel Insights into Transposon Genes and Non-Coding RNAs during Atlantic Salmon Seawater Adaptation.

Simple SummaryThis study proposes a novel approach to analyze transcriptome data sets using the Atlantic salmon seawater adaptation process as a model. Salmon salar smolts were transferred to seawater under two strategies: (i) fish group exposed to gradual salinity changes (GSC) and (ii) fish group exposed to a salinity shock (SS). mRNA and miRNAs sequencing were performed for gills, intestine, and head kidney tissues. The whole-genome transcript expression profiling revealed specific gene expression patterns among the tissues and treatments. A great abundance of transposable elements was observed in chromosome regions differentially expressed under experimental conditions. Moreover, small RNA expression analysis suggested fewer of miRNAs associated with the smoltification process. However, target analysis of these miRNAs suggests a regulatory role of process such as growth, stress response, and immunity. The findings uncover whole-transcriptome modulation during seawater adaptation of Atlantic salmon, evidencing the interplaying among mRNAs and miRNAs.The growing amount of genome information and transcriptomes data available allows for a better understanding of biological processes. However, analysis of complex transcriptomic experimental designs involving different conditions, tissues, or times is relevant. This study proposes a novel approach to analyze complex data sets combining transcriptomes and miRNAs at the chromosome-level genome. Atlantic salmon smolts were transferred to seawater under two strategies: (i) fish group exposed to gradual salinity changes (GSC) and (ii) fish group exposed to a salinity shock (SS). Gills, intestine, and head kidney samples were used for total RNA extraction, followed by mRNA and small RNA illumina sequencing. Different expression patterns among the tissues and treatments were observed through a whole-genome transcriptomic approach. Chromosome regions highly expressed between experimental conditions included a great abundance of transposable elements. In addition, differential expression analysis showed a greater number of transcripts modulated in response to SS in gills and head kidney. miRNA expression analysis suggested a small number of miRNAs involved in the smoltification process. However, target analysis of these miRNAs showed a regulatory role in growth, stress response, and immunity. This study is the first to evidence the interplaying among mRNAs and miRNAs and the structural relationship at the genome level during Atlantic salmon smoltification.

Microanatomy of the antennal glands and their Na+/K+-ATPase activity in three true crab species (Brachyura), Portunus pelagicus (Linnaeus, 1758) (Portunidae), Macrophthalmus dentipes Lucas in Guérin, 1836 (Macrophthalmidae) and Eriocheir hepuensis Dai, 1991 (Varunidae)

Abstract There are differences between various crab species in the function and structure of organs involved in ionic and osmotic regulation processes. The antennal glands together constitute one of the most important organs involved in the osmoregulation in crabs. The present investigation aimed to study the tissue structure of the antennal glands and their Na+/K+-ATPase (NKA) pump activity in three true crab species from three different habitats, including the marine (Portunus pelagicus (Linnaeus, 1758)), the estuarine (Macrophthalmus dentipes Lucas in Guérin, 1836) and the freshwater habitat (Eriocheir hepuensis Dai, 1991). In this regard, the tissue structure of the antennal glands and the activity of the Na+/K+-ATPase (NKA) pump were assessed in these three selected species. The results showed that the antennal glands in all studied species consisted of two anterior parts and a posterior part. The anterior parts are composed of the proximal tubular region (PT) and the distal tubular region (DT). The PT and DT parts comprised the coelomosac and labyrinths in the anterior portion, and the bladder located in the posterior portion. However, despite the similarity in the general tissue structure of the antennal gland in marine, estuarine and freshwater crab species, some structural differences were observed between those species. Labyrinth cells, coelomosac podocytes, and bladder cells in the estuarine crab M. dentipes contained large vacuoles especially on the top (i.e., near the lumen) of the cells. The highest amount of NKA pump activity was measured in the antennal glands of M. dentipes (). The NKA pump plays a more important role in the estuary and seawater adaptation of crabs, but freshwater species are not highly dependent on the NKA enzyme for osmoregulation.

Effects of constant and diel cyclic temperatures on the liver and intestinal phospholipid fatty acid composition in rainbow trout Oncorhynchus mykiss during seawater acclimation

BackgroundRainbow trout is an economically important fish in aquaculture and is a model species in environmental physiology. Despite earlier research on the seawater adaptability of rainbow trout at different temperature regimes, the influence on the liver and intestine in this species is still unknown. Two trials were conducted to investigate the effects of constant and diel cyclic temperatures on phospholipid fatty acid (PLFA) composition in the liver and intestine of rainbow trout during seawater acclimation.ResultsAt the end of growth trial 1, fish at 9 and 12.5 °C showed significantly higher ratios of unsaturated to saturated (U/S) and unsaturation index (UI) than those at 16 °C in liver and intestine phospholipids. After day 1 of seawater acclimation, the U/S, UI, and average chain length (ACL) of liver and intestinal phospholipids in fish at 16 °C significantly increased. Two weeks after seawater acclimation, the liver and intestinal PLFA composition adapted to salinity changes. In trial 2, significantly higher U/S, UI, and ACL were found in intestinal phospholipids at 13 ± 2 °C. On the first day after seawater acclimation, UI and ACL in liver phospholipids significantly increased at 13 °C, while fish at 13 ± 2 °C showed significantly decreased U/S, UI, and ACL in the intestine. At the end of growth trial 2, liver PLFA compositions were stable, whereas intestinal PLFA at 13 and 13 ± 1 °C showed significantly decreased U/S, UI, and ACL. A two-way analysis of variance and principal component analysis revealed significant effects of different constant temperatures, seawater acclimation, and their interaction on the liver and intestinal phospholipids, a significant effect of diel cyclic temperature on intestinal phospholipids, and the effects of seawater acclimation and its interaction with diel cyclic temperature on liver phospholipids.ConclusionTemperatures of 9 and 12.5 °C could elevate membrane fluidity and thickness in the liver and intestine of rainbow trout in freshwater, whereas no significant effects were found with diel temperature variations. After seawater acclimation, constant and diel cyclic temperatures significantly influenced the membrane fluidity and thickness of the liver and intestine. Compared with constant temperature, diel temperature variation (13 ± 2 °C) can enhance the adaptability of rainbow trout during seawater acclimation.

Transcriptome Profiling of Long Non-coding RNAs During the Atlantic Salmon Smoltification Process.

For salmon aquaculture, one of the most critical phase is the parr-smolt transformation. Studies around this process have mainly focused on physiological changes and the Na+/K+-ATPase activity during the osmoregulatory activity. However, understanding how the salmon genome regulates the parr-smolt transformation, specifically the molecular mechanisms involved, remains uncovered. This study aimed to explore the transcriptional modulation of long non-coding RNAs (lncRNAs), as key molecular regulators, during the freshwater (FW) to seawater (SW) transfer in Atlantic salmon. Transcriptome sequencing was performed from gill samples of Atlantic salmon adapted from FW to SW through gradual salinity changes from 0 to 30 PSU. The results showed that most transcripts differently modulated were downregulated in all salinity conditions. Relevant biological processes were associated with growth, collagen formation, immune response, metabolism, and heme transport. Notably, 2864 putative lncRNAs were identified in Atlantic salmon gills differently expressed during fish smoltification. The highest number of lncRNAs differently modulated was observed at 30 PSU. Correlation expression analysis suggests putative regulatory roles of lncRNAs with smoltification-related genes. Herein, co-localization of Na+/K+-ATPase, growth hormone receptor, and thyroid hormone receptor genes with lncRNAs differentially expressed suggest putative regulatory mechanisms in the Atlantic salmon genome. The lncRNAs can be used as novel biomarkers for the fish smoltification process. Here, the lncRNA_145326 and lncRNA_18762 are putatively related to the parr-smolt transfer in Atlantic salmon. This study is the first description of lncRNAs with putative regulatory roles in Atlantic salmon during the SW adaptation.

A comparison of two seawater adaptation strategies for Atlantic salmon post-smolt (Salmo salar) grown in recirculating aquaculture systems (RAS): Nitrification, water and gut microbiota, and performance of fish

The land-based production phase in Norwegian Atlantic salmon farming has the past years been extended to include post-smolt for an increasing number of farmers. Post-smolt production can involve introduction of brackish/seawater to a recirculating aquaculture system (RAS) that is acclimatized to freshwater for the earlier stages of salmon production. A change from freshwater to seawater shifts the environmental conditions for fish, bacteria and water treatment processes in RAS. Two regimes for salinity increase were studied to evaluate the effects on nitrification functionality, water and gut microbiota and fish performance on land and in the sea cages. A fish group of 200,000 salmon parr were stocked in a brackish water RAS (bRAS) at 3‰ salinity. After the fish had smoltified the group was split in two, one group was kept in bRAS and the other was moved to a RAS operated at 28‰ salinity (sRAS). The bRAS was operated with a gradual increase in salinity from 3 to 26‰ over a period of 28 days, whereafter both groups were moved to two separate sea cages. Bacterial communities of water, biofilter biofilm and fish faeces were characterized by 16S rRNA amplicon sequencing. Nitrification capacity tests at different salinities were performed on biofilter media from bRAS, to evaluate short term robustness to salinity changes. Ordination based on Bray-Curtis similarities showed that in water samples in bRAS, the bacterial communities were stable from 12 to 26‰ salinity increase. The faecal microbiota of the fish showed high inter-individual variation within fish tanks, suggesting stochastic processes/drift to affect the community structures in addition to salinity increase. The same nitrifying bacteria were present in bRAS (throughout the salinity increase) and in sRAS, showing that these nitrifiers could adapt to salinities from 3 to 26‰, and 28‰. After the sea cage phase, fish from the sRAS system had in total 2.9% higher weight than the fish from bRAS, however the mortality was 15% higher in the sRAS group. Salinity was a driver for succession in RAS, and other factors such as organic load in the water and stochastic processes in the host also affected the bacterial community dynamics.

Differential Seawater Adaptability in Three Different Sizes of Under-yearling Steelhead Trout.

Seawater adaptability of steelhead trout increases along with the increase in the size of the fish, independent of parr-smolt transformation. Three 96 h seawater challenge tests were conducted to determine the size at which seawater adaptability of steelhead trout develops. Plasma Na+ and Cl- levels, moisture content, gill Na+/K+ ATPase activity, and mortality during the 96 h after direct transfer to seawater (32 ppt) were determined. Plasma Na+ and Cl- levels in 50 g fish continuously increased during the 96 h after the transfer to seawater (p<0.05), but the levels in 100 and 150 g fish leveled off after 24 h (p<0.05). Both 100 and 150 g size steelhead trout maintained muscle moisture content (%) better than 50 g size fish (p<0.05). Gill Na+/K+ ATPase activity in the 100 g size group increased in a time-dependent manner after transfer to seawater (p<0.05), whereas activity in the 50 and 150 g sizes did not increase (p>0.05), for which a possible explanation was discussed. A mere 2.6% mortality in both the 50 and 150 g size groups was observed. In conclusion, the current results indicate that 50 g size steelhead trout did not show development of a high level of hypoosmoregulatory capacity, whereas fish in the 100 and 150 g size groups showed a high level in our experimental conditions. Therefore, the steelhead trout larger than a 100 g size is recommended for transfer to seawater culture.

The vertebrate Aqp14 water channel is a neuropeptide-regulated polytransporter

Water channels (aquaporins) were originally discovered in mammals with fourteen subfamilies now identified (AQP0-13). Here we show that a functional Aqp14 subfamily phylogenetically related to AQP4-type channels exists in all vertebrate lineages except hagfishes and eutherian mammals. In contrast to the water-selective classical aquaporins, which have four aromatic-arginine constriction residues, Aqp14 proteins present five non-aromatic constriction residues and facilitate the permeation of water, urea, ammonia, H2O2 and glycerol. Immunocytochemical assays suggest that Aqp14 channels play important osmoregulatory roles in piscine seawater adaptation. Our data indicate that Aqp14 intracellular trafficking is tightly regulated by the vasotocinergic/isotocinergic neuropeptide and receptor systems, whereby protein kinase C and A transduction pathways phosphorylate highly conserved C-terminal residues to control channel plasma membrane insertion. The neuropeptide regulation of Aqp14 channels thus predates the vasotocin/vasopressin regulation of AQP2-5-6 orthologs observed in tetrapods. These findings demonstrate that vertebrate Aqp14 channels represent an ancient subfamily of neuropeptide-regulated polytransporters.

Effects of different photoperiod regimes on the smoltification and seawater adaptation of seawater-farmed rainbow trout (Oncorhynchus mykiss): Insights from Na+, K+–ATPase activity and transcription of osmoregulation and growth regulation genes

Photoperiod is thought to be the main zeitgeber that induces smoltification in salmonids. However, its effects on the smoltification of rainbow trout (Oncorhynchus mykiss) are not fully understood and no published data documents the effects of the photoperiod regime currently used commercially, continuous light (LL). The present study compared the effect of four different photoperiod regimes (i.e. advanced phase photoperiod (APP), delayed phase photoperiod (DPP), LL and simulated natural photoperiod (SNP)) on the smoltification and growth of juvenile rainbow trout during their freshwater phase of winter-spring and the following summer post smolt phase. Smoltification was evaluated by monitoring gill Na+,K+–ATPase (NKA) activity and transcription of NKA α-subunit isoforms 1a and 1b, and Na+,K+,2Cl‐cotransporter 1a. Growth was measured as specific growth rate of both length and weight, and through molecular growth proxies such as the levels of circulating insulin-like growth factor 1 (IGF-I) in plasma and transcription of igf-I, igf binding protein 1b (igfbp1b), growth hormone receptor 1 (ghr1) and cathepsin L (ctsl) in the liver. Results indicate that APP induces a longer smolt window and higher levels of plasma IGF-I in both freshwater and seawater (two months post transfer), while DPP led to a shorter smolt window, lower plasma IGF-I levels in freshwater and seawater, an earlier decrease in liver igf-I and ctsl transcription in freshwater (as seen by modelling over time) and lower specific growth rate in freshwater. The transcription analysis of osmoregulatory genes complemented NKA activity and allowed for the detection of a transient response to light and of differences between the osmoregulatory capacity of parr and desmolted fish. Furthermore, an upregulation of the liver transcription of igf-I, ghr1 and ctsl was found in all treatments during the smolt window, which corresponded to the periods with highest growth. Finally, both plasma IGF-I and liver igf-I in seawater were found to be significantly correlated to fish growth in seawater. However, our data did not show that plasma IGF-I prior to seawater transfer could be used as a reliable predictor of growth in seawater. Overall, and especially when compared with other salmonid species, photoperiod seems to be a weaker inducer of smoltification in rainbow trout, according to the parameters that were tested, suggesting that other environmental cues might be more important drivers of this process.

Do the toll-like receptors and complement systems play equally important roles in freshwater adapted Dolly Varden char (Salvelinus malma)?

Unlike the normal anadromous lifestyle, Chinese native Dolly Varden char (Salvelinus malma) is locked in land and lives in fresh water lifetime. To explore the effect of freshwater adaption on its immune system, we constructed a pooled cDNA library of hepatopancreas and spleen of Chinese freshwater Dolly Varden char (S. malma). A total of 27,829 unigenes were generated from 31,233 high-quality transcripts and 17,670 complete open reading frames (ORF) were identified. Totally 25,809 unigenes were successfully annotated and it classified more native than adaptive immunity-associated genes, and more genes involved in toll-like receptor signal pathway than those in complement and coagulation cascades (51 vs 3), implying the relative more important role of toll-like receptors than the complement system under bacterial injection for the freshwater Dolly Varden char. These huge different numbers of TLR and complement system identified in freshwater Dolly Varden char probably caused by distinct evolution pressure patterns between fish TLR and complement system, representative by TLR3 and TLR5 as well as C4 and C6, respectively, which were under purifying and positively selecting pressure, respectively. Further seawater adaptation experiment and the comparison study with our library will no doubt be helpful to elucidate the effect of freshwater adaption of Chinese native Dolly Varden char on its immune system.

The Amphibious Mudskipper: A Unique Model Bridging the Gap of Central Actions of Osmoregulatory Hormones Between Terrestrial and Aquatic Vertebrates.

Body fluid regulation, or osmoregulation, continues to be a major topic in comparative physiology, and teleost fishes have been the subject of intensive research. Great progress has been made in understanding the osmoregulatory mechanisms including drinking behavior in teleosts and mammals. Mudskipper gobies can bridge the gap from aquatic to terrestrial habitats by their amphibious behavior, but the studies are yet emerging. In this review, we introduce this unique teleost as a model to study osmoregulatory behaviors, particularly amphibious behaviors regulated by the central action of hormones. Regarding drinking behavior of mammals, a thirst sensation is aroused by angiotensin II (Ang II) through direct actions on the forebrain circumventricular structures, which predominantly motivates them to search for water and take it into the mouth for drinking. By contrast, aquatic teleosts can drink water that is constantly present in their mouth only by reflex swallowing, and Ang II induces swallowing by acting on the hindbrain circumventricular organ without inducing thirst. In mudskippers, however, through the loss of buccal water by swallowing, which appears to induce buccal drying on land, Ang II motivates these fishes to move to water for drinking. Thus, mudskippers revealed a unique thirst regulation by sensory detection in the buccal cavity. In addition, the neurohypophysial hormones, isotocin (IT) and vasotocin (VT), promote migration to water via IT receptors in mudskippers. VT is also dipsogenic and the neurons in the forebrain may mediate their thirst. VT regulates social behaviors as well as osmoregulation. The VT-induced migration appears to be a submissive response of subordinate mudskippers to escape from competitive and dehydrating land. Together with implications of VT in aggression, mudskippers may bridge the multiple functions of neurohypophysial hormones. Interestingly, cortisol, an important hormone for seawater adaptation and stress response in teleosts, also stimulates the migration toward water, mediated possibly via the mineralocorticoid receptor. The corticosteroid system that is responsive to external stressors can accelerate emergence of migration to alternative habitats. In this review, we suggest this unique teleost as an important model to deepen insights into the behavioral roles of these hormones in relation to osmoregulation.