Producing a larger post‐smolt in recirculating aquaculture systems (RAS) could shorten the production time in sea cages and potentially reduce mortality. Knowledge of the biological requirements of ...post‐smolts in closed‐containment systems is however lacking. In the present study, the effects of salinity and water velocity on growth, survival, health, and welfare of Atlantic salmon reared in RAS were examined. Salmon smolts were stocked in three separate RAS with salinities of 12, 22, and 32‰ and subjected to high (1.0 body lengths per s−1) or low (0.3 body lengths second−1) water velocity. Growth performance, survival, welfare, and physiological stress responses were monitored until the fish reached a bodyweight of around 450 g. Growth rate was higher at lower salinity and higher water velocity generally had a positive effect on growth in all salinities. Feed conversion ratio was lower at 12‰ compared to the 22 and 32‰ when the fish were between 250 and 450 g. Higher mortality, elevated plasma cortisol levels, higher incidence of cataract, and a higher expression of stress‐induced genes in the skin (iNOS, Muc5ac‐like) indicated a negative effect of higher salinity on fish welfare. Male maturation was low (<1%), and not affected by salinity or water velocity.
The sea-run phenotype of rainbow trout (Oncorhynchus mykiss), like other anadromous salmonids, present a juvenile stage fully adapted to life in freshwater known as parr. Development in freshwater is ...followed by the smolt stage, where preadaptations needed for seawater life are developed making fish ready to migrate to the ocean, after which event they become post-smolts. While these three life stages have been studied using a variety of approaches, proteomics has never been used for such purpose. The present study characterised the blood plasma proteome of parr, smolt and post-smolt rainbow trout using a gel electrophoresis liquid chromatography tandem mass spectrometry approach alone or in combination with low-abundant protein enrichment technology (combinatorial peptide ligand library). In total, 1,822 proteins were quantified, 17.95% of them being detected only in plasma post enrichment. Across all life stages, the most abundant proteins were ankyrin-2, DNA primase large subunit, actin, serum albumin, apolipoproteins, hemoglobin subunits, hemopexin-like proteins and complement C3. When comparing the different life stages, 17 proteins involved in mechanisms to cope with hyperosmotic stress and retinal changes, as well as the downregulation of nonessential processes in smolts, were significantly different between parr and smolt samples. On the other hand, 11 proteins related to increased growth in post-smolts, and also related to coping with hyperosmotic stress and to retinal changes, were significantly different between smolt and post-smolt samples. Overall, this study presents a series of proteins with the potential to complement current seawater-readiness assessment tests in rainbow trout, which can be measured non-lethally in an easily accessible biofluid. Furthermore, this study represents a first in-depth characterisation of the rainbow trout blood plasma proteome, having considered three life stages of the fish and used both fractionation alone or in combination with enrichment methods to increase protein detection.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Environmental challenges related to open sea cage production of Atlantic salmon have sparked interest in developing commercial-scale semi-closed sea systems for post-smolt Atlantic salmon (100–1000 ...g). Determining the mass-specific water flow required by post-smolts will largely influence the design and dimensioning of such systems. In this experiment, post-smolts were exposed to four levels of specific water flow: 0.2, 0.3, 0.4, and 0.5 L kg fish−1 min−1. All treatments involved flow-through seawater with full oxygenation, a salinity of 34‰, and a mean temperature of 9.3 °C. The stocking density was kept stable at 75 kg m−3. Water pH decreased with reduced flow, while partial pressure of carbon dioxide (pCO2) and total ammonia nitrogen (TAN) in the water increased. The increase in water CO2 was reflected in the blood with increased pCO2, HCO3−, and decreased Cl− in the lowest water flow treatment (0.2 L kg fish−1 min−1), indicating a typical regulatory response to increased water CO2 over the eight-week experimental period. No negative effects on osmoregulation, external macroscopic welfare, or performance indicators were observed, suggesting that within the time period of this experiment, post-smolts can compensate for reductions in water flow down to 0.2 L kg fish−1 min−1. However, to avoid activating and exhausting potentially energy-costly physiological regulatory mechanisms, it is suggested to keep specific water flow above 0.3 L kg fish−1 min−1 in large-scale operations with semi-closed sea systems at intermediate temperatures.
Rainbow trout (Oncorhynchus mykiss) is a salmonid species with a complex life-history. Wild populations are naturally divided into freshwater residents and sea-run migrants. Migrants undergo an ...energy-demanding adaptation for life in seawater, known as smoltification, while freshwater residents display these changes in an attenuated magnitude and rate. Despite this, in seawater rainbow trout farming all fish are transferred to seawater. Under these circumstances, weeks after seawater transfer, a significant portion of the fish die (around 10%) or experience growth stunting (GS; around 10%), which represents an important profitability and welfare issue. The underlying causes leading to GS in seawater-transferred rainbow trout remain unknown. In this study, we aimed at characterising the GS phenotype in seawater-transferred rainbow trout using untargeted and targeted approaches. To this end, the liver proteome (LC-MS/MS) and lipidome (LC-MS) of GS and fast-growing phenotypes were profiled to identify molecules and processes that are characteristic of the GS phenotype. Moreover, the transcription, abundance or activity of key proteins and hormones related to osmoregulation (Gill Na+, K + -ATPase activity), growth (plasma IGF-I, and liver igf1, igfbp1b, ghr1 and ctsl) and stress (plasma cortisol) were measured using targeted approaches. No differences in Gill Na+, K + -ATPase activity and plasma cortisol were detected between the two groups. However, a significant downregulation in plasma IGF-I and liver igf1 transcription pointed at this growth factor as an important pathomechanism for GS. Changes in the liver proteome revealed reactive-oxygen-species-mediated endoplasmic reticulum stress as a key mechanism underlying the GS phenotype. From the lipidomic analysis, key observations include a reduction in triacylglycerols and elevated amounts of cardiolipins, a characteristic lipid class associated with oxidative stress, in GS phenotype. While the triggers to the activation of endoplasmic reticulum stress are still unknown, data from this study point towards a nutritional deficiency as an underlying driver of this phenotype.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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.
•High temperature (18 °C) was the main factor stimulating early sexual maturation.•The effect of restricted feed ration on early maturation was temperature-dependent.•Onset of maturation seemed triggered by the photoperiod switch from short to long-light.•Signs of smoltification were obvious primarily at 8 °C and only in immature salmon.•Use of sustained high temperature in salmon aquaculture can increase risk of early maturation.
Maturation of Atlantic salmon male postsmolts is a concern in aquaculture due to its increasing occurrence under intensive rearing conditions and its negative impact on growth, welfare and seawater ...readiness. The effect of temperature and photoperiod on maturation was assessed in male postsmolts kept in freshwater. We used a 2 × 2 factorial design with two temperatures (12.5 and 15 °C) and two photoperiods (a group in continuous light LD24:0 or LL, and another receiving a 5-week LD12:12 winter signal or WS). Salmon in the four resulting treatments (1000 parr, initial mean weight 52.1 ± 5.2 g) were reared in a flow-through system from 28 October 2019 to 30 May 2020. Morphology (body weight, condition factor), maturation indicators including gonadosomatic index (GSI), plasma 11-Ketotestosterone (11-KT), pituitary follicle-stimulating hormone β-subunit (fshb) and luteinizing hormone β-subunit (lhb) transcript levels, as well as smoltification markers (Na+, K+- ATPase activity) were assessed. Results revealed that rearing salmon at 15 °C was the most important factor promoting early maturation, leading to 100% of males maturing in late May in 15-WS, and 75% in 15-LL. However, the groups receiving a winter signal (WS) displayed a highly synchronized onset and progression of maturation specially at 15 °C, revealed by the low variability observed among individuals in GSI and fshb transcription after the WS. This evidences the role of the photoperiod switch from short to long day as zeitgeber for sexual maturation. On the contrary, under constant light (LL), entry into maturation was not synchronized among individuals, and onset of maturation occurred spontaneously in a proportion of males highly dependent upon temperature (75% in 15-LL, 25% in 12.5-LL). Signs of smoltification were poor at both temperatures, and the WS did not induce development of hypo-osmoregulatory abilities in any case. This suggests that a winter signal may not induce smoltification if introduced at high temperature or when fish have reached large size, and instead may increase the risk of a sexual maturation response. These findings are relevant for the aquaculture industry, since similar rearing conditions are currently used in the industry, including constant high water temperature and winter signal regimes. The use of such conditions can increase the risk of early maturation, as well as of poor hypo-osmoregulatory performance.
•High temperature (15 °C) induced early sexual maturation of males under both photoperiods.•A switch in photoperiod from short to long-light synchronized onset of maturation.•Smoltification did not occur in response to the change in photoperiod.•Combining high temperatures and photoperiod cues increases risk of early maturation.
Sexual maturation of post-smolts is a concern for Atlantic salmon producers, and its occurrence is dependent upon factors such as water temperature and energy availability, among others. The present ...study was designed to investigate the effect of different temperatures and feeding regimes on testis development and local regulation of spermatogenesis in salmon post-smolts. A 3 × 2 factorial design was used, with three temperatures (8 °C, 12.5 °C, and 18 °C) and two feed regimes (100% and 67% ration). Salmon (1800 parr, initial mean weight 23.1 ± 7.2 g) were reared from 28 October 2018 to 30 May 2019 in a freshwater flow-through system under continuous light (LD24:0), except for a 5-week winter signal (LD12:12) introduced on 4 February 2019. Testis histology, transcription of follicle-stimulating hormone receptor (fshr) and luteinizing hormone receptor (lhr), and transcription of testis proteins involved in spermatogenesis regulation such as gonadal-soma-derived factors 1 (gsdf1) and 2 (gsdf2), anti-Müllerian hormone (amh), and insulin-like growth factor-3 (igf3), were analyzed. Results showed that high temperature alone (18 °C), irrespective of the feed regime, promoted early presence of type B spermatogonia and reduced transcript levels of the proliferation-inhibitory factor amh in males still considered immature, of groups 18–100% and 18–67% that later matured in high proportion (~100%). This effect was also present to some degree in the group 12.5–100% (40% maturation), and absent in 12.5–67%, 8–100%, and 8–67% (groups with little or no maturation). Later, at onset of rapid testis growth, high temperature was linked to a pronounced downregulation of amh and of the self-renewal factors gsdf1 and gsdf2, as well as to a pronounced upregulation of the proliferation-stimulating factor igf3. Overall, the present findings demonstrate that rearing salmon at high temperatures can stimulate an early activation of the brain–pituitary–gonad axis before actual onset of rapid testis growth, enhancing and accelerating the mechanisms that control initiation and progression of maturation, while the feed regime has a minor impact. This poses a challenge for current salmon aquaculture practices that use sustained high temperature to maximize growth, since these practices increase the risk of early post-smolt maturation.
The use of closed containment (CCS) or semi‐closed containment systems (S‐CCS) for Atlantic salmon Salmo salar aquaculture is under evaluation in Norway. One such system is the Preline S‐CCS, a ...floating raceway system that pumps water from 35 m depth creating a constant current through the system. Exposing fish to moderate water currents is considered aerobic exercise and it is often perceived as positive for fish welfare, growth, food utilization, muscle development and cardiac health. The present study compared fish reared in the Preline S‐CCS and in a reference open pen. Samples were taken in fresh water before being transferred to the seawater systems and after 1, 2 and 4 months in seawater and analysed for growth, mortality, muscle development and plasma insulin‐like growth factor I (IGF‐I) levels. Moreover, gene transcription were determined in the skeletal muscle igf‐I, insulin‐like growth factor 1 receptor a (igf1ra) and insulin‐like growth factor 1 binding protein 1a (igf1bp1a) and cardiac transcription factors myocyte‐specific enhancer factor 2C (mef2c), gata4 and vascular endothelial growth factor (vegf). While the results suggest that post‐smolts in Preline S‐CCS were smaller than reference fish, fish from Preline S‐CCS have less accumulated mortality at the end of the experiment and showed 2.44 times more small muscle fibres than the reference group fish after 4 months in seawater. These results confirmed what was previously observed in the second generation of Preline. Similar levels of big muscle fibres between Preline S‐CCS and reference suggest a similar hypertrophy of muscle fibres even with lower IGF‐I expression in the Preline S‐CCS. Cardiac gene transcription suggests cardiac hypertrophy was observed after 4 months in seawater in the Preline S‐CCS group. Altogether, Preline S‐CCS is a promising technology able to produce more robust S. salar with a faster growth and lower mortality in the subsequent standard open cage system growth period.
Smoltification in salmonids occurs during spring in response to increasing photoperiod to prepare for marine life. Smoltification is associated with increased hypo-osmoregulatory ability and enhanced ...growth potential, mediated by growth hormone and insulin-like growth factor (IGF)-1. Rainbow trout is uniquely insensitive to the induction of smoltification-associated changes by photoperiod, such as the activation of gill Na+,K+-ATPase (NKA). We measured the circulating IGF-1 and IGF-binding protein (IGFBP)-2b levels in yearling rainbow trout exposed to natural and manipulated photoperiods during spring and correlated these with gill NKA activity and body size. Although the effect of photoperiod manipulation on body size and circulating IGF-1 and IGFBP-2b was negligible, they were positively correlated with gill NKA activity in fish under simulated natural photoperiod. We next pit-tagged yearling rainbow trout and fed them a restricted ration or to satiation under a natural photoperiod. In April, gill NKA activity was higher in the satiation group than in the restricted group and positively correlated with body size and growth rate. In addition, circulating IGFBP-2b was positively correlated with gill NKA, size and growth, whereas circulating IGF-1 was correlated only with size and growth. The relationship between circulating IGF-1 and growth intensified from May to June, suggesting that the IGF-1–growth relationship was disrupted in April when gill NKA was activated. Two additional IGFBPs were related to growth parameters but not to gill NKA activity. The present study suggests that circulating IGFBP-2b and IGF-1 mediate the size-dependent activation of gill NKA in yearling rainbow trout during spring.
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•Profiles of circulating IGF-1 and IGFBP-2b in yearling rainbow trout in spring were established.•Gill Na+,K+-ATPase activity was correlated with body size, IGF-1 and IGFBP-2b levels.•Feeding manipulation affected gill Na+,K+-ATPase activity in spring.•Circulating IGFBP-2b may be involved in the activation of gill Na+,K+-ATPase in trout.
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.
•nkaα1a, nkaα1b and nkcc1a provide relevant information that the NKA activity does not reflect.•The liver transcription of igf-I, ghr1 and ctsl is directly related to fish SGRL.•While the best photoperiod in terms of duration of the smolt window and abundance of plasma IGF-I seemed to be APP, DPP was clearly the worst one.•While plasma IGF-I and liver igf-I in seawater was correlated to growth, plasma IGF-I prior seawater transfer was not a predictor of growth.
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