Seawater Tolerance Research Articles

New wine in old bottles: Modification of the Na+/K+‐ATPase enzyme activity assay and its application in salmonid aquaculture

AbstractThe Na+, K+ ATPase (NKA) enzyme is important to generate the transmembrane ion gradient in the gills, intestine, and kidneys, hence, is vital for secondary transport of fluids and different solutes in teleosts. Gill NKA enzyme activity is often used as a proxy for parr–smolt transformation (PST) during which anadromous salmonids prepare for seawater (SW). Increased intensification and production of larger smolts in modern salmonid aquaculture has resulted in reports of gill NKA activity being less reliable as a proxy for smolt quality. Consequently, changes in mRNA nka‐α1b/α1a ratios in gills are increasingly used as indicators of PST. However, nka isoform mRNA abundance may not reflect translation into the functional protein, nor the activity of the mature enzyme. This may limit the predictive power of molecular markers under certain environmental conditions, rearing regimes and biological scenarios. During PST, the osmoregulatory transformations necessary for SW tolerance and survival does not only occur in the gills. Equally important are the changes in ion transporting activities, including NKA activity, in the intestine and kidneys. However, to our knowledge, there are no previous studies addressing the timing and concurrent changes in NKA activity in the three osmoregulatory tissue during PST. Here we present modifications and optimization of the NKA enzyme activity protocols for gill, intestinal and kidney tissue and outline how to best utilize NKA activity measurements as part of a more holistic approach to evaluate overall smolt quality in modern aquaculture.

The GH/IGF axis in the sea lamprey during metamorphosis and seawater acclimation

How the growth hormone (GH)/insulin-like growth factor (IGF) system affects osmoregulation in basal vertebrates remains unknown. We examined changes in the expression of components of the GH/IGF axis and gill ion transporters during metamorphosis and following seawater (SW) exposure of sea lamprey. During metamorphosis, increases in gill nka and nkcc1 and salinity tolerance were accompanied by increases in pituitary gh, liver igf1, gill ghr and igf1, but not liver ghr. SW exposure of fully metamorphosed sea lamprey resulted in slight increases in plasma chloride concentrations after SW exposure, indicating a high level of SW tolerance, but no major changes in mRNA levels of gill ion transporters or components of the GH/IGF axis. Our results indicate that metamorphosis is a critical point in the lifecycle of sea lamprey for stimulation of the GH/IGF axis and is temporally associated with and likely promotes metamorphosis and SW tolerance.

Effect of Photoperiod and Transfer Time on Atlantic Salmon Smolt Quality and Growth in Freshwater and Seawater Aquaculture Systems

Smoltification is a key process in Atlantic salmon aquaculture, given it prepares the fish for a successful transit from fresh to seawater. However, industry players have not yet reached a consensus on the best protocols to produce high-quality smolts. In this study, we assessed how the combination of two photoperiod regimes in freshwater (continuous light or LL, and natural photoperiod or LDN) and four transfer times to seawater (February, March, April, and May) affected smolt development and their subsequent growth in seawater until slaughter during commercial production. The results demonstrated that smoltification and growth in freshwater were only slightly modulated by the photoperiod treatment and were instead much more affected by the limiting effect of the low water temperature during that period. In seawater, the growth rate was the highest in the same groups, which had, however, experienced a delay in growth when in freshwater, and consequently, no differences in the final body weight between the eight treatments were found. Such compensatory growth in the sea was probably enhanced by the increasing smolt quality, which could allow for better performance in seawater. A significant link between the weight at slaughter and weight at transfer was observed only in the groups with a lower smolt quality (LL-Feb, LDN-Feb and LDN-Mar), which suggests that larger individuals could cope better with a saline environment. In contrast, smaller smolts probably suffered greater osmotic stress that hindered their performance at sea. Afterwards, as smolt quality increased in the subsequent transfer groups, the relevance of this size effect decreased. This means that the industry may benefit from transferring larger smolts to seawater, especially if these are suspected of having developed suboptimal seawater tolerance. Those individuals are likely to cope better with saline conditions than smaller smolts. Future research should focus on the possible long-term effects of freshwater-rearing regimes on smolt performance in the seawater phase.

Long term effects of smolt production strategy and early seawater phase rearing environment on mortality, growth, sexual maturation, and vertebra deformities in farmed Atlantic salmon (Salmo salar L.)

The life history of Atlantic salmon is plastic and determined by factors such as daylength and temperature. In aquaculture, artificial long days during smolt production maximize growth and allows for early sea-transfer, occurring between the first summer and autumn after hatching. However, the impact of such intensive rearing strategies on the salmon's welfare is not well understood. In this study, we follow underyearling (0+) juvenile salmon through two different smolt production regimes, either dietary induced seawater tolerance (fed with SuperSmolt®) under continuous light (LLS), or light-induced seawater tolerance, where for six weeks the photoperiod was lowered to 12 h light and 12 h darkness and then increased again to continuous light (LD-LL). Next, the LD-LL and LLS salmon were distributed into indoor seawater tanks where they experienced light and temperature conditions simulating either an August (AUG) or an October (OCT) sea transfer scenario lasting 2 months, after which all fish were transferred into a sea cage and reared there until reaching harvest size (10 months). Welfare parameters were mortality, growth, vertebra deformities (radiology), and sexual maturation. LLS grew faster than LD-LL smolts up until seawater transfer, while LD-LL post-smolts grew faster during the early seawater phase, resulting in equal harvest weights. The simulated AUG transfer gave higher harvest weights than the simulated OCT transfer regardless of smolt production strategy. Mortality during the first period in sea cages was higher in the LD-LL compared to LLS fish within the AUG scenario. At harvest, male sexual maturation was significantly higher in the LLS AUG (21%) group compared to the other groups (∼10%), while the occurrence of fish with vertebra deformities (1 ≥ deformed vertebrae) was highest among LD-LL fish within the AUG scenario (31%). Only the LD-LL fish had severely deformed individuals (fish with >10 deformed vertebrae): 2.8% in AUG and 5.6% in OCT. The present study shows that the incidence of male sexual maturation and vertebral deformities is lower in dietary- and photoperiod-induced smolts, respectively, by transferring them to seawater in autumn rather than summer. However, such a strategy could be expected to reduce the harvest weight of both LD-LL- and LLS-induced smolts.

Growth responses and genetic variation among highly ecologically diverse spring wheat genotypes grown under seawater stress.

Most of the freshwaters worldwide are used for agriculture. Freshwater sources are expected to decline and will not suffice to support the food production needed for the growing population. Therefore, growing crops with seawater might constitute a solution. However, very little work has been done on the effect of seawater stress on wheat, an important cereal crop. The present study aimed to determine whether particular wheat genotypes provided better resistance to seawater stress. A set of 80 highly diverse spring wheat genotypes collected from different countries in Europe, Asia, Africa, North and South America was exposed to 50% seawater stress at the early growth stage. Four seeding shoot and root traits were scored for all genotypes. High genetic variations were found among all genotypes for the epicotyl length (EL), hypocotyl length (HL), number of radicles (NOR), and fresh weight (FW). Eight genotypes with high-performance scores of seedling traits were selected. The correlation analyses revealed highly significant correlations among all traits scored in this study. The strongest correlation was found between the NOR and the other seeding traits. Thus, the NOR might be an important adaptive trait for seawater tolerance. The genetic diversity among all genotypes was investigated based on genetic distance. A wide range of genetic distances among all genotypes was found. There was also a great genetic distance among the eight selected genotypes. In particular, the genetic distance between ATRI 5310 (France) and the other seven genotypes was the greatest. Such high genetic diversity might be utilized to select highly divergent genotypes for crossing in a future breeding program. The present study provides very useful information on the presence of different genetic resources in wheat for seawater tolerance.

The importance of seawater tolerance and native status in mediating the distribution of inland fishes

AbstractAimUnravelling the ecological and historical factors that underlie species distributions has challenged ecologists for a long time. Thus, our objective is to understand the role of environmental variables explaining the distribution of three major eco‐evolutionary groups of inland fishes (Darlington's divisions: primary, i.e. strict freshwater; secondary, i.e. salt‐tolerant; and peripheral, i.e. diadromous and marine origin), and how these variables are related to fish traits.LocationIberian Peninsula.Taxon51 native and 17 alien inland fish species from the Iberian Peninsula.MethodsWe modelled distributions of the most common inland fish species across the Iberian Peninsula to compare the importance of different predictors among the three Darlington's divisions and between native and alien species. To explore the importance of specific environmental variables in determining the distribution of different traits of inland fish, variable importances obtained from species distribution models were subjected to a redundancy analysis.ResultsDarlington's divisions differ significantly in salinity tolerance, in distribution overlap, in the importance of distribution predictors and associated life‐history traits. Topographic and climatic variables were generally more important than land use and anthropogenic factors in explaining fish distributions. We found significant differences in the importance of variables explaining the distribution of native vs. alien species and especially among Darlington's divisions. River basin was most important for primary native and many alien species. Increasing mean temperature and damming were positively associated with the presence of tolerant, large‐bodied and warm‐water alien species from more hydrologically stable habitats.Main conclusionsDespite marked differences in the distribution patterns of native and alien species, evolutionary and introduction histories as well as seawater tolerance are central factors explaining the current distribution of inland fishes. Darlington's divisions proved useful for addressing ecological and biogeographical questions at broader spatial scales.

Evaluating acid-aluminum stress in streams of the Northeastern U.S. at watershed, fish community and physiological scales

In spite of overall improvements in air and water quality, biological stress from low pH and high concentrations of inorganic aluminum continue to impact fish and fish habitat in northeastern North America, with independent and interactive effects on individuals, populations and communities. Integrative indicators can therefore be useful in monitoring both impact and recovery across multiple scales. Using coupled water chemistry (pH, conductivity, and base cation and inorganic aluminum concentration), geographic (site elevation and watershed area) and biological (fish diversity, fish abundance, gill aluminum concentration and gill physiology) data, we developed an integrated indicator of acid-aluminum stress across the White and Green mountains in central New England, USA. As has been established in a number of previous studies, preliminary analysis clearly indicated that across all sites, inorganic aluminum concentration was consistently greatest during the spring season. Structural Equation modeling (SEM) revealed that toxic conditions (concurrent low pH and high concentrations of inorganic aluminum) were well summarized with an integrated toxicity score, related to both base cation concentrations and elevation, with sites at higher elevations more likely to experience toxic conditions as well as low base cation concentrations. Fish diversity and abundance generally trended negatively with toxicity score, with fewer cyprinids and sculpins at high toxicity score sites. In spite of considerable variation among individuals, gill aluminum was positively related to toxicity score for both Atlantic salmon and brook trout. Observed elevated gill aluminum levels associated with reduced gill metabolic activity in Atlantic salmon smolts from impacted systems likely result in impaired osmoregulatory function and seawater tolerance. Overall, our results suggest that the integrated toxicity score metric is associated with a syndrome of acute physiological stress, reduced abundance, and low species diversity for sensitive stream fishes in New England, and can likely serve as a reliable indicator of continued impairment or recovery of acid-aluminum vulnerable systems in this ecoregion.

Juvenile sea lamprey (Petromyzon marinus) have a wide window of elevated salinity tolerance that is eventually limited during springtime warming

The present study examined changes in biometric characteristics, osmoregulatory capacity, and seawater (SW) tolerance of juvenile sea lamprey ( Petromyzon marinus) throughout the varying thermal changes from late autumn to late spring. Body length, mass, and condition factor were maintained until April, when significant declines in mass and condition factor were observed to correspond with increases in temperature. Nearly 100% survival in SW was maintained through April. In May, after river and estuarine temperatures had increased, significant mortality in SW (up to 50%) was observed. After SW acclimation, plasma chloride was maintained at an elevated set point, and gill Na+/K+-ATPase activity was elevated. Neither parameter appeared to be affected during springtime warming. Together, these results provide a first characterization of the sustained osmoregulatory performance of juvenile sea lamprey after metamorphosis and show that the window of increased hypo-osmoregulatory performance for SW entry lasts for at least 5 months but may ultimately be limited by increases in river water temperatures in late spring.

Na+/HCO3- cotransporter 1 (nbce1) isoform gene expression during smoltification and seawater acclimation of Atlantic salmon.

The life history of Atlantic salmon (Salmo salar) includes an initial freshwater phase (parr) that precedes a springtime migration to marine environments as smolts. The development of osmoregulatory systems that will ultimately support the survival of juveniles upon entry into marine habitats is a key aspect of smoltification. While the acquisition of seawater tolerance in all euryhaline species demands the concerted activity of specific ion pumps, transporters, and channels, the contributions of Na+/HCO3- cotransporter 1 (Nbce1) to salinity acclimation remain unresolved. Here, we investigated the branchial and intestinal expression of three Na+/HCO3- cotransporter 1 isoforms, denoted nbce1.1, -1.2a, and -1.2b. Given the proposed role of Nbce1 in supporting the absorption of environmental Na+ by ionocytes, we first hypothesized that expression of a branchial nbce1 transcript (nbce1.2a) would be attenuated in salmon undergoing smoltification and following seawater exposure. In two separate years, we observed spring increases in branchial Na+/K+-ATPase activity, Na+/K+/2Cl- cotransporter 1, and cystic fibrosis transmembrane regulator 1 expression characteristic of smoltification, whereas there were no attendant changes in nbce1.2a expression. Nonetheless, branchial nbce1.2a levels were reduced in parr and smolts within 2days of seawater exposure. In the intestine, gene transcript abundance for nbce1.1 increased from spring to summer in the anterior intestine, but not in the posterior intestine or pyloric caeca, and nbce1.1 and -1.2b expression in the intestine showed season-dependent transcriptional regulation by seawater exposure. Collectively, our data indicate that tissue-specific modulation of all three nbce1 isoforms underlies adaptive responses to seawater.

Network analysis reveals that acute stress exacerbates gene regulatory responses of the gill to seawater in Atlantic salmon.

The transition from freshwater to seawater represents a physiological challenge for Atlantic salmon smolts preparing for downstream migration. Stressors occurring during downstream migration to the ocean impair the ability of smolts to maintain osmotic/ionic homeostasis in seawater. The molecular mechanisms underlying this interaction are not fully understood, especially at the organ level. We combined RNA-Seq with measures of whole-animal homeostasis to examine gene expression dynamics in the gills of smolts associated with impaired seawater tolerance after an aquaculture-related stressor. Smolts were given a 24 h seawater tolerance test before and after exposure to an acute handling/confinement stress. RNA-Seq followed by differential expression and weighted gene correlation network analysis (WGCNA) was used to quantify the transcriptional response of the gill to handling/confinement stress, seawater and their interaction. Exposure to acute stress was associated with a general stress response and impaired osmotic/ionic homeostasis in seawater. We identified gene networks in the gill exhibiting response to acute stress alone, seawater alone, and others exhibiting combined effects of both stress and seawater. Our findings indicate that acute handling/confinement stress increases the intensity of seawater-related gene expression and suggest that increased investment in mechanisms related to ion transport may be part of a compensatory response to impaired seawater tolerance in smolts.

Diluted Seawater and Ammonia-N Tolerance of Two Mangrove Crab Species. New Insights to Understand the Vulnerability of Pristine Islands Ecosystems Organisms

Mangrove ecosystems are the primary receptors of anthropogenic pollution in tropical areas. Assessing the vulnerability of these ecosystems can be expressed, among other indicators, by studying the health of ‘ecosystem engineers’. In this study, mangrove forests facing opposing anthropogenic pressures were studied (i) in the uninhabited island of Europa (Mozambique Channel), considered as a pristine ecosystem, and, (ii) on the island of Mayotte, facing regular domestic wastewater discharges. Using an ecophysiological approach, the effects of diluted seawater (DSW) and increased ammonia-N were studied for two fiddler crab species: Gelasimus tetragonon (GT) on the island of Europa and Paraleptuca chlorophthalmus (PC) on the island of Mayotte. Osmoregulation curves and osmoregulatory capacity were determined along with O2 consumption rates after a 96 h exposure period. Histological analyses were also carried out on two important metabolic organs: the hepatopancreas and the posterior gills. Results indicate that both crab species are good hyper-hypo-osmoregulators but only PC can maintain its osmoregulatory capacity when exposed to ammonia-N. Oxygen consumption is increased in GT after 96 h of exposure to ammonia-N but this does not occur in PC. Finally, a thickening of the gill osmoregulatory epithelium was observed after 96 h in PC when exposed to ammonium but not in GT. Therefore, the two species do not have the same tolerance to DSW and increased ammonia-N. PC shows physiological acclimation capacities in order to better manage nitrogenous enrichments. GT did not show the same physiological plasticity when exposed to ammonia-N and could be more at risk by this kind of stress. These results along with those from other studies regarding the effects of domestic effluents on mangrove crabs are discussed. Therefore, the greater vulnerability of organisms occupying pristine ecosystems could induce major changes in mangrove functioning if crabs, that are engineer species of the ecosystem, are about to reduce their bioturbation activity or, even, disappear from the mangrove forests.

Corticosteroid control of Na+/K+-ATPase in the intestine of the sea lamprey (Petromyzon marinus)

Anadromous sea lamprey (Petromyzon marinus) larvae undergo a months-long true metamorphosis during which they develop seawater (SW) tolerance prior to downstream migration and SW entry. We have previously shown that intestinal Na+/K+-ATPase (NKA) activity increases during metamorphosis and is critical to the osmoregulatory function of the intestine in SW. The present study investigated the role of 11-deoxycortisol (S) in controlling NKA in the anterior (AI) and posterior (PI) intestine during sea lamprey metamorphosis. In a tissue profile, nka mRNA and protein were most abundant in the gill, kidney, and AI. During metamorphosis, AI nka mRNA increased 10-fold, whereas PI nka mRNA did not change. Specific corticosteroid receptors were found in the AI, which had a higher binding affinity for S compared to 11-deoxycorticosterone (DOC). In vivo administration of S in mid-metamorphic lamprey upregulated NKA activity 3-fold in the AI and PI, whereas administration of DOC did not affect intestinal NKA activity. During a 24 h SW challenge test, dehydration of white muscle moisture was rescued by prior treatment with S, which was associated with increased intestinal nka mRNA and NKA activity. These results indicate that intestinal osmoregulation in sea lamprey is a target for control by S during metamorphosis and the development of SW tolerance.

Cortisol is an osmoregulatory and glucose-regulating hormone in Atlantic sturgeon, a basal ray-finned fish.

Our current understanding of the hormonal control of ion regulation in aquatic vertebrates comes primarily from studies on teleost fishes, with relatively little information on more basal fishes. We investigated the role of cortisol in regulating seawater tolerance and its underlying mechanisms in an anadromous chondrostean, the Atlantic sturgeon (Acipenser oxyrinchus). Exposure of freshwater-reared Atlantic sturgeon to seawater (25 ppt) resulted in transient (1-3 day) increases in plasma chloride, cortisol and glucose levels and long-term (6-14 day) increases in the abundance of gill Na+/K+/2Cl- cotransporter (NKCC), which plays a critical role in salt secretion in teleosts. The abundance of gill V-type H+-ATPase, which is thought to play a role in ion uptake in fishes, decreased after exposure to seawater. Gill Na+/K+-ATPase activity did not increase in 25 ppt seawater, but did increase in fish gradually acclimated to 30 ppt. Treatment of Atlantic sturgeon in freshwater with exogenous cortisol resulted in dose-dependent increases in cortisol, glucose and gill NKCC and H+-ATPase abundance. Our results indicate that cortisol has an important role in regulating mechanisms for ion secretion and uptake in sturgeon and provide support for the hypothesis that control of osmoregulation and glucose by corticosteroids is a basal trait of jawed vertebrates.

Cortisol regulates insulin-like growth-factor binding protein (igfbp) gene expression in Atlantic salmon parr

The growth hormone (Gh)/insulin-like growth-factor (Igf)/Igf binding protein (Igfbp) system regulates growth and osmoregulation in salmonid fishes, but how this system interacts with other endocrine systems is largely unknown. Given the well-documented consequences of mounting a glucocorticoid stress response on growth, we hypothesized that cortisol inhibits anabolic processes by modulating the expression of hepatic igfbp mRNAs. Atlantic salmon (Salmo salar) parr were implanted intraperitoneally with cortisol implants (0, 10, and 40 μg g−1 body weight) and sampled after 3 or 14 days. Cortisol elicited a dose-dependent reduction in specific growth rate (SGR) after 14 days. While plasma Gh and Igf1 levels were unchanged, hepatic igf1 mRNA was diminished and hepatic igfbp1b1 and -1b2 were stimulated by the high cortisol dose. Plasma Igf1 was positively correlated with SGR at 14 days. Hepatic gh receptor (ghr), igfbp1a, -2a, -2b1, and -2b2 levels were not impacted by cortisol. Muscle igf2, but not igf1 or ghr, levels were stimulated at 3 days by the high cortisol dose. As both cortisol and the Gh/Igf axis promote seawater (SW) tolerance, and particular igfbps respond to SW exposure, we also assessed whether cortisol coordinates the expression of branchial igfbps and genes associated with ion transport. Cortisol stimulated branchial igfbp5b2 levels in parallel with Na+/K+-ATPase (NKA) activity and nka-α1b, Na+/K+/2Cl--cotransporter 1 (nkcc1), and cystic fibrosis transmembrane regulator 1 (cftr1) mRNA levels. The collective results indicate that cortisol modulates the growth of juvenile salmon via the regulation of hepatic igfbp1s whereas no clear links between cortisol and branchial igfbps previously shown to be salinity-responsive could be established.

11-Deoxycortisol controls hydromineral balance in the most basal osmoregulating vertebrate, sea lamprey (Petromyzon marinus)

It is unknown whether and how osmoregulation is controlled by corticosteroid signaling in the phylogenetically basal vertebrate group Agnatha, including lampreys and hagfishes. It is known that a truncated steroid biosynthetic pathway in lampreys produces two predominant circulating corticosteroids, 11-deoxycortisol (S) and 11-deoxycorticosterone (DOC). Furthermore, lampreys express only a single, ancestral corticosteroid receptor (CR). Whether S and/or DOC interact with the CR to control osmoregulation in lampreys is still unknown. We examined the role of the endogenous corticosteroids in vivo and ex vivo in sea lamprey (Petromyzon marinus) during the critical metamorphic period during which sea lamprey increase osmoregulatory capacity and acquire seawater (SW) tolerance. We demonstrate in vivo that increases in circulating [S] and gill CR abundance are associated with increases in osmoregulatory capacity during metamorphosis. We further show that in vivo and ex vivo treatment with S increases activity and expression of gill active ion transporters and improves SW tolerance, and that only S (and not DOC) has regulatory control over active ion transport in the gills. Lastly, we show that the lamprey CR expresses an ancestral, spironolactone-as-agonist structural motif and that spironolactone treatment in vivo increases osmoregulatory capacity. Together, these results demonstrate that S is an osmoregulatory hormone in lamprey and that receptor-mediated discriminative corticosteroid regulation of hydromineral balance is an evolutionarily basal trait among vertebrates.

Unexpectedly long survivorship on seawater of multiple coastal beetles indicates the possibility of “floating dispersal” for transoceanic migration

AbstractOcean is generally a major barrier of migration for most flightless land animals; however, terrestrial flightless insects often demonstrate wide range distribution across oceans. To elucidate the mechanism of flightless insects to expand their distribution over the sea, we measured the survivorship of nine species of coastal beetles on seawater (i.e. seawater tolerance). We observed that two out of nine coastal beetle species showed over 10 days of median survival days (50% death days), and nearly 1 month of maximum survival days. From the unexpectedly long survival on the surface of seawater, we propose the hypothesis of “floating dispersal,” where individuals simply float on the sea surface and cross the ocean with currents. This mode can provide a novel explanation of transoceanic migration/distribution of flightless animals.

Diminished Expression of Na+/HCO3−cotransporter 1.2a (nbce1.2a) mRNA Accompanies Seawater Acclimation in Atlantic Salmon

The life history of anadromous Atlantic salmon (Salmo salar) entails an initial freshwater stage that precedes their migration to marine environments. The development of physiological systems that will ultimately support the survival of juvenile salmon upon their entry into marine habitats is termed parr‐smolt transformation. This springtime transformation (smoltification) includes the acquisition of seawater (SW) tolerance. Thus, the ability of smolts to successfully acclimate to SW is attained through the development of branchial pathways for ion‐secretion. In addition, the capacity to rapidly attenuate ion‐absorptive processes upon exposure to SW is a vital aspect of SW tolerance. In the current study, we investigated the branchial gene expression patterns of three Na+/HCO3−cotransporter 1 isoforms (nbce1.1, nbce1.2a, and nbce1.2b) during smoltification and following seawater challenges. Given the proposed roles of Nbce1s in the absorption of environmental Na+ by branchial ionocytes in other teleosts, we hypothesized that nbce1s would be downregulated in salmon undergoing smoltification and/or following SW exposure. In two separate years, we observed springtime increases in branchial Na+/K+‐ATPase activity, Na+/K+/2Cl−‐cotransporter 1 and cystic fibrosis transmembrane regulator 1 expression characteristic of smoltification, whereas there were no seasonal changes in the expression of nbce1 isoforms. On the other hand, we observed diminished nbce1.2a expression within 2 days after the transfer of parr and smolts to SW that remained low throughout the experiment (>2 weeks). Collectively, our results suggest that while there are no clear seasonal nbce1 dynamics during smoltification, the modulation of nbce1.2a expression upon exposure to SW may underlie the attenuation of environmental Na+ uptake.Support or Funding InformationSupported by the National Science Foundation (IOS‐1755131).

Intestinal Gene Expression of Na+/HCO3− cotransporter 1 (nbce1) Isoforms in Atlantic Salmon During Smoltification and Seawater Acclimation

In preparation for migration from freshwater to marine habitats, Atlantic salmon undergo smoltification, a springtime transformation that includes the development of seawater tolerance. The ability of smolts to inhabit seawater is attained through the coordinated development of ion and water‐transporting processes in key organs such as the gill, kidney, and intestine. To promote solute‐linked water absorption, the intestinal epithelium of fishes acclimated to SW secretes HCO3− to form luminal Ca2+ and Mg2+ precipitates. As a source of HCO3− for apical anion exchange, Na+/HCO3− cotransporter 1 (Nbce1) facilitates the transport of plasma HCO3− into the interior of epithelial cells across the basolateral membrane. While Atlantic salmon (Salmo salar) express three nbce1 isoforms (nbce1.1, nbce1.2a, and nbce1.2b), there is no information on whether their expression corresponds with the development of intestinal fluid absorption that occurs during smoltification. We first determined that nbce1.1 and nbce1.2b gene transcripts were highly expressed in pyloric cacae and anterior intestine of smolts, whereas nbce1.2a showed low expression in the intestine but robust expression in gill. Next, we sampled salmon undergoing parr‐smolt transformation and observed elevated branchial Na+/K+‐ATPase activity in May characteristic of smoltification. Moreover, smolts exhibited an improved ability to maintain ionoregulatory balance when transferred to seawater in May versus March. We will report corresponding patterns of nbce1.1 and nbce1.2b expression in pyloric cacae and anterior intestine during smoltification and following seawater transfer to resolve whether their transcriptional patterns correspond with the acquisition of intestinal fluid absorptive processes.Support or Funding InformationSupported by the National Science Foundation (IOS‐1755131).

Muscle Protease Activities in Salmo Trutta L. Inhabiting the Krivoi Ruchey River

The effects of size, age, and smoltification on muscle protein breakdown systems were studied in wild brown trout Salmo trutta L. inhabiting the Krivoi Ruchey River of the White Sea basin, Kola Peninsula. Activities of autophagy-related proteases including cathepsin B and D, calpains, and proteasome were assayed in the skeletal muscle of brown trout parr and smolt of different age group. Youngest fish group consisted of the most actively growing individuals possessed the higher rate of protein breakdown compared to older groups. Different patterns of muscle protein breakdown inherent to brown trout parr and smolts were shown to be associated with calpain system and cathepsin D activities. Thus, increased activity of these proteases in smoltifying individuals obviously results in amino acid accumulation that could be a mechanism of seawater tolerance required for seaward migration. This study is the first to show the age- and stage-related dynamics in protease activities in skeletal muscle of brown trout inhabiting the Krivoi Ruchey River. Growth- and smoltification-related patterns of protease activities were quite similar in brown trout from the Krivoi Ruchey River and previously studied rivers of the White Sea basin, however, some habitat-related differences were observed.

Impact of environmental factors on the growth and maturation of farmed Arctic charr

AbstractFarming of Arctic charr mainly takes place in land‐based farms applying intensive rearing methods with relatively high production costs. Depending on local conditions at each site, it is possible to regulate important environmental factors to improve productivity and well‐being of the fish. Knowledge about how these different environmental factors affect various farming traits is important to reduce production costs. This review shows how rearing temperature, photoperiod, salinity and feeding rate can affect farming traits such as growth rate, maturation and feed conversion efficiency of Arctic charr. High growth rate during juvenile phase when the fish are reared at higher temperatures can result in higher incidence of maturation during the on‐growing period. Overall, more moderate rearing temperature regimes seem to result in better long‐term growth rates. Photoperiod manipulation and feed ration can be used as tools to improve growth and reduce maturation. It is possible to rear Arctic charr successfully up to market size in salinities up to c. 27–28 ppt. However, extended rearing resulted in higher ratio of sexually mature fish at 29 than at 25 possibly linked to higher rearing temperatures in brackish water. Future studies should focus on better preserving the potential high growth rate of Arctic charr during juvenile phase into the on‐growing period and establish protocols to improve the seawater tolerance of Arctic charr.