Weeks In Freshwater Research Articles

In silico, in vitro and in vivo characterization of host-associated Latilactobacillus curvatus strains for potential probiotic applications in farmed Atlantic salmon (Salmo salar)

Salmon aquaculture is the fastest growing animal protein production system in the world; however, intensive farming leads to poor weight gain, stress, and disease outbreaks. Probiotics offer the potential to enhance growth performance and feed efficiency in Atlantic salmon, as well as immunostimulate fish against common pathogens, benefitting farmers and consumers with more efficient production. Here, we isolated and identified 900 native microbial isolates including 18 Lactobacilli from the farmed salmon intestines. Based on whole-genome sequencing and phylogenetic analysis, the Lactobacillus candidates belonged to Latilactobacillus curvatus (L. curvatus) species and formed two distinct phylogenetic groups. Using bioinformatics and in vitro analyses, we selected two candidates L. curvatus ATCC PTA-127116 and L. curvatus ATCC PTA-127117, which showed desirable safety and probiotic properties. The two L. curvatus candidates were evaluated for safety and efficacy (higher final weight) in Atlantic salmon alongside spore-forming Bacilli isolated from salmon, poultry, and swine. All the tested candidates were safe to salmon with no adverse effects. While we did not see efficacy in any Bacillus supplemented groups, compared to untreated group, the group administered with the two L. curvatus strains consortium in feed for seven weeks in freshwater showed indicators of improvement in final body weight by 4.2%. Similarly, the two L. curvatus candidates were also evaluated for safety and efficacy in Atlantic salmon in saltwater; the group administered with the two L. curvatus strains consortium in feed for 11 weeks showed indicators of improvement in final body weight by 4.7%. Comprehensive metabolomics analyses in the presence of different prebiotics and/or additives identified galactooligosaccharide as a potential prebiotic to enhance the efficacy of two L. curvatus candidates. All together, these data provide comprehensive genomic, phenotypic and metabolomic evidence of safety and desirable probiotic properties as well as indicators of in vivo efficacy of two novel endogenous L. curvatus candidates for potential probiotic applications in Atlantic salmon. The in vivo findings need to be confirmed in larger performance studies, including field trials.

Impact of dietary zinc and seawater transfer on zinc status, availability, endogenous loss and osmoregulatory responses in Atlantic salmon smolt fed low fish meal feeds

Atlantic salmon fed low fish meal feeds supplemented with zinc (Zn) were studied in two feeding trials. In trial I, Atlantic salmon parr were fed six graded Zn levels (40 to 249 mg kg−1 as ZnSO4) for 8 weeks in freshwater followed by a 4-week seawater phase. In trial II, Atlantic salmon post-smolt were fed for 10 weeks in SW with 10 dietary Zn levels (45 to 280 mg kg−1), either as ZnSO4 or Zn-glycinate. Growth was unaffected by dietary Zn in both trials. Dietary Zn affected concentration of Na + and K+ ions in plasma, branchial and intestinal expression of sodium potassium ATPase, tissue and body Zn status, and cataracts. Seawater transfer significantly reduced apparent availability, body and tissue levels of Zn due to increased endogenous Zn loss. Atlantic salmon post-smolt in seawater improved body and tissue Zn status with increasing dietary Zn levels, irrespective of the Zn source. Body or tissue saturation of Zn occurred at dietary Zn levels between 137 and 156 mg kg−1 with smolts in freshwater and 181 to 218 mg kg−1 in SW post-smolts. Dietary Zn levels below 180 mg kg−1 in low fish meal feeds compromised the Zn status and welfare of Atlantic salmon in seawater.

Dietary electrolyte balance of Atlantic salmon (Salmo salar) freshwater feeds: Impact on osmoregulation, mineral metabolism and performance in seawater

Dietary electrolyte balance is the equilibrium of monovalent cations and anions that influence the acid-base balance of the feed (dEB = Na + K − Cl, mEq kg−1). Dietary electrolytes/minerals can influence the physiological changes during smoltification in Atlantic salmon. In this context, we aimed to study if the dEB of the freshwater feeds can be used to pre-adapt the hypoosmotic functionality and the associated effects on mineral metabolism. The dEB of commercial freshwater Atlantic salmon feeds in Norway varied from −9 to 400 mEq kg−1 feed. Three experimental feeds were formulated to study incremental levels of dEB reflecting the low (L-dEB, −50 to 0), median (M-dEB, 200–250) and high (H-dEB, 350–400). Triplicate groups of Atlantic salmon parr (36 g) were fed one of the three feeds for 8 weeks in freshwater at 12 °C. The fish were transferred to full strength seawater in indoor tanks and fed a commercial diet for 6 weeks. Growth was not differentially affected by dEB levels, neither in the freshwater phase nor in the seawater. Plasma electrolytes (Na+ and Cl−) and gill mRNA expression of sodium potassium ATPase (NKA a1b, seawater isoform) were significantly lower in L-dEB fed fish. In the intestine, carbonate precipitates 24 h after seawater transfer was higher in fish fed both L-dEB and H-dEB feeds compared to the M-dEB fed fish. Whole body and plasma mineral levels were significantly affected by dEB levels in freshwater feeds. Interestingly, the carryover effect of dEB in freshwater feeds was significant after 6 weeks in seawater for plasma and whole-body Zn status, with the H-dEB fed fish showing significantly increased body Zn status compared to L-dEB and M-dEB fed fish. The study revealed that mineral metabolism and intestinal response to seawater transfer can be pre-adapted by modulating the electrolyte and/or mineral balance in freshwater feeds in Atlantic salmon. Further, dEB did not affect long term development of cataract or vertebral deformities.

The transcriptomic response of adult salmon lice (Lepeophtheirus salmonis) to reduced salinity

Salmon lice (Lepeophtheirus salmonis) are marine parasitic copepods living on salmonids and are challenging for salmon aquaculture. One of several treatment methods is the application of freshwater to the fish which can lead to lice loss. However, lab experiments have shown that salmon lice, acclimated to seawater, are capable of surviving for several weeks in freshwater, when attached to a host. If not attached to a host, they die within a few hours in freshwater but can survive a longer time in brackish water. The molecular mechanisms involved in the adaptation to low salinity of the louse have not been identified yet. In this study we incubated salmon lice, being attached to a host, or detached, in seawater, brackish water and freshwater for 4 h and 1 d, sampled the animals and used RNA-Seq to identify genes involved in these mechanisms. Freshwater incubation led to a much stronger regulatory response than brackish water and a longer incubation time gave a stronger effect than a short incubation. Among the most interesting genes, upregulated in low salinity water are in addition to several transporters, several enzymes involved in amino acid metabolism and especially in the proline biosynthesis. A strong upregulation of these enzymes might lead to an accumulation of proline which is known to be used as an osmolyte in other species. While the RNA-Seq experiment was performed with female samples, qPCR showed that at least 10 genes regulated in females, were also regulated in males.

Re-evaluating the dietary requirement of EPA and DHA for Atlantic salmon in freshwater

The requirement for eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) is recommended at 0.50–1.0% of the dry diet for juvenile Atlantic salmon (Salmo salar). Current commercial aquafeed formulations for Atlantic salmon tend to contain a physiological excess of these fatty acids. The aim of this study was to re-evaluate the EPA and DHA requirement by Atlantic salmon parr in freshwater since it was determined 20 years ago, and feed formulations and salmon production have changed. This study evaluated the effect of different dietary levels of EPA and DHA (0.25 to 2.0%) on growth performance, overall health, and whole-body EPA and DHA content. Salmon (initial weight 22.6 g ± 1.2) were fed four levels (0.25, 0.5, 0.75 and 2%) of EPA + DHA diets for 6 weeks in freshwater at 12 °C. Diets were fed to triplicate tanks, with 20 fish per tank. There were no mortalities throughout the trial and fish nearly doubled their weight after 6 weeks. Categorically, there were no discrete significant differences in growth performance among treatments after 6 weeks. However, weight gain was exponentially related to diet EPA + DHA levels (p < .0001; r2 = 28.3%), and the feed conversion ratio was inversely related. In both cases, these relationships suggested that the asymptote is reached between 0.25% and 0.5% dietary EPA + DHA. Whole body DHA and EPA showed positive linear relationships with diet DHA (p < .001; r2 = 88.7%) and EPA (p < .001; r2 = 92.9%). Based on growth performance and whole-body fatty acid content, salmon fed a diet containing 0.50% EPA + DHA diet exhibited similar growth performance and haematocrit with control diet (2%). Therefore, 0.5% is considered the lowest required EPA + DHA level for Atlantic salmon parr in freshwater in this study.

Atlantic salmon fed a nutrient package of surplus methionine, vitamin B12, folic acid and vitamin B6 improved growth and reduced the relative liver size, but when in excess growth reduced

Feeding plant‐based diet through smoltification of Atlantic salmon requires verification of the optimal level of 1C nutrients. Here, we fed Atlantic salmon plant‐based diets containing three different surplus amounts of the 1C nutrients; methionine, cobalamin (vitamin B12), pyridoxine (vitamin B6) and folic acid during 6 weeks in fresh water, through smoltification, followed by 3 months on‐growing period in salt water. The three diets were fed to fish dispersed in triplicate tanks throughout the experiment. Mean start body weight was 32 g. Dietary methionine levels in the diets were 6.7, 9.2 and 11.7 g/kg. Dietary B6 was 6.75, 8.45 and 11 mg/kg. Cobalamin was 0.16, 0.18 and 0.20 mg/kg. While dietary folic acid was 2.9, 4.8 and 6.3 mg/kg, diets are referred to as low, medium and high 1C diet. All other amino acids were similar between diets. The results showed no differences in growth or feed utilization in the fresh water period, but following the on‐growing salt water period, differences between diets occurred. The fish fed the medium 1C diet showed better growth, as compared to fish fed the low or high 1C diet (p = .009). The medium 1C fed fish showed a relative lower liver weight compared with fish fed low or high 1C diet (p = .025). Condition factor was better in fish fed the medium and high 1C diet as compared to those fed the low 1C diet (p = .0006). As expected, free methionine in liver, plasma and muscle increased by dietary methionine inclusion. Surplus vitamins only had minor effect on tissue concentrations. Based on these findings, we conclude that the micronutrient and methionine level presented in the medium 1C diet improved the growth, liver size and condition factor; however, more research is needed to evaluate the optimal requirement level for each of the 1C nutrients.

Influence of Dietary Supplementation of Probiotic Pediococcus acidilactici MA18/5M During the Transition From Freshwater to Seawater on Intestinal Health and Microbiota of Atlantic Salmon (Salmo salar L.).

The aim of this study was to assess the effect of the transfer from freshwater to seawater on the distal intestinal bacterial communities of Atlantic salmon (Salmo salar L.) and to evaluate the effect of dietary inclusion of Pediococcus acidilactici MA18/5M (at 1.19 × 106 CFU/g). In this context, fish health and antiviral response were also investigated. A 12-week feeding trial was conducted in a flow-through rearing system involving 6 weeks in freshwater and 6 weeks in seawater. Fish received a control and probiotic diet. The composition of the salmon gut bacterial communities was determined by high-throughput sequencing of digesta and mucosa samples from both the freshwater and seawater stage. The main phyla detected during both freshwater and seawater stages were Firmicutes, Proteobacteria, Fusobacteria, and Actinobacteria. Significant differences were observed between the intestinal microbiota in the digesta and the mucosa. Both probiotic supplementation and the seawater transfer (SWT) had a substantial impact on the microbial communities, with most pronounced changes detected in the mucosal communities after SWT. This last finding together with a significantly higher antiviral response (mx-1 and tlr3 gene expression) in the distal intestine of fish fed the probiotic diet suggest a causal link between the microbiota modulation and activation of antiviral response. Feeding probiotics during the freshwater stage did not significantly increase survival after infectious pancreatic necrosis virus (IPNV) challenge after SWT, although higher survival was observed in one out of two replicate challenge tanks. In conclusion, this study demonstrated that both dietary probiotic supplementation and transfer from freshwater to seawater have an important role in modulating the bacterial communities in the distal intestine of Atlantic salmon. Furthermore, supplementation of the diet with P. acidilactici MA18/5M can modulate antiviral response.

Interactive effects of dietary magnesium and vitamin E on growth performance, body composition, blood parameters and antioxidant status in Japanese seabass (Lateolabrax japonicus) fed oxidized oil

A study was conducted to investigate effects and interactions of magnesium (Mg) and vitamin E (VE) on growth performance, body composition, hepatic antioxidant capacity and serum biochemistry parameters of juvenile Japanese seabass Lateolabrax japonicus under oxidative stress condition. Fish (initial average body weight of 6.10 ± 0.20 g) were fed 9 oxidized oil diets supplemented with 3 grade levels of Mg (0, 520 and 1570 mg kg−1 diet) and VE (0, 60 and 200 mg kg−1 diet) for 8 weeks in freshwater. The results showed that diets supplemented 520 mg kg−1 Mg and/or 60 mg kg−1 VE had highest growth and muscle lipid content. There were highest total superoxide dismutase, catalase, glutathione peroxidase activities and lowest malondialdehyde content in liver of fish fed diets supplemented 520 mg kg−1 Mg and/or 60 mg kg−1 VE. Contrary to Mg concentrations, Ca concentrations and Ca/Mg ratio in whole body were inversely related to dietary Mg levels. However, combined deficiency or excess of dietary Mg and VE led to the decrease of hepatic antioxidant capacity, body lipid deposition and growth of Japanese seabass under oxidative stress condition.

Fresh water acclimation elicits a decrease in plasma corticosteroids in the euryhaline Atlantic stingray, Dasyatis sabina

It is thought that the elasmobranch corticosteroid hormone 1α-hydroxycorticosterone (1α-B) functions as both a glucocorticoid (GC) and mineralocorticoid (MC). Classical antinatriuretic MC activities would run counter to the osmoregulatory strategy of euryhaline elasmobranchs acclimating to fresh water (FW). Therefore we hypothesize that FW acclimation will be accompanied by a decrease in plasma corticosteroids in these animals. However, events that activate the “fight-or-flight” response could mask changes associated with acclimation to lower salinities. To better define the MC role of corticosteroids in elasmobranchs, we designed a transfer system that allows the acclimation of Atlantic stingrays (Dasyatis sabina) from seawater (SW) to FW over 12h while minimizing other extraneous stressors. Blood and interrenal glands were sampled from one group of stingrays 24h after FW transfer, while another group was sampled two weeks after FW transfer. Two other groups served as mock-transfer controls in that they were treated and sampled in the same way, but remained in SW for the entire period. Plasma corticosteroids, osmolality, chloride, and urea were significantly lower in FW-acclimated stingrays (compared to mock-transfer stingrays) 24h after FW transfer. This pattern remained after two weeks in FW, with the exception that plasma corticosteroids returned to pre-acclimation levels. There were no significant differences between experimental groups in interrenal levels of mRNAs encoding key steroidogenic proteins (steroidogenic acute regulatory protein and cholesterol side chain cleavage enzyme). Temporally decreased corticosteroid levels during FW acclimation are consistent with the unique strategy of euryhaline elasmobranchs, whereby lower plasma osmolality is maintained in FW vs. SW environments to reduce hydromineral gradients.

Use of camelina oil to replace fish oil in diets for farmed salmonids and Atlantic cod

Abstract In this study growth and lipid composition of Atlantic salmon, rainbow trout and Atlantic cod were compared after feeding a diet containing camelina oil (CO) as a full replacement of fish oil (FO). CO is an interesting candidate for replacement due to its high total lipid content (40%) and a high amount of the ω3 precursor, 18:3ω3; high levels of mono- and polyunsaturated fatty acids, as well as significant amounts of γ-tocopherol, a potent antioxidant. Diets were formulated to meet the nutritional requirements for each species, and either contained FO or CO as the main lipid source. Three separate experiments were conducted (triplicate tanks for FO and CO): Atlantic salmon (mean initial weight 242 ± 46 g) were fed experimental diets for 16 weeks in seawater at 14 °C, rainbow trout (44.9 ± 10 g) were fed for 12 weeks in freshwater at 14 °C, and Atlantic cod (14.4 ± 1.6 g) were fed for 13 weeks at 10 °C. Growth performance of Atlantic cod was significantly affected by dietary CO in comparison to cod fed the FO diet (p

The effect of water temperature on vertebral deformities and vaccine-induced abdominal lesions in Atlantic salmon, Salmo salar L.

This study investigates the effects of water temperature (T) on vaccine-induced abdominal lesions (i.p. injection with oil-adjuvant vaccine) and vertebral deformities in Atlantic salmon. Quadruple groups of vaccinated (V) or unvaccinated (U) underyearling smolts were reared in tanks under four different temperature regimes for 6 weeks in fresh water (FW) followed by 6 weeks in sea water (SW). The four different T regimes were 10 °C FW-10 °C SW (10-10), 10 °C FW-16 °C SW (10-16), 16 °C FW-10 °C SW (16-10) and 16 °C FW-16 °C SW (16-16). After the temperature regimes were finished, the fish were group-tagged and transferred to a common sea cage for on-growth until harvest size. At termination, weight was significantly affected by both T and V, while lesion score and deformities were affected by T only. The weight difference between the largest and smallest U group was 20.3% (16-10 U: 2.4 kg, 10-16 U: 1.89 kg), while the largest difference between U and V fish within a T regime was 28.7% (16-16 U: 2.1 kg, 16-16 V: 1.5 kg). Fish from the 16-16, 16-10 and 10-16 regimes had a significant higher lesion score than those from the 10-10 regime. Fish from the 10-16 and 16-16 regimes displayed a significantly higher prevalence of vertebral deformities (palpation : 13-27%, radiology: 88-94%) than fish from the 10-10 and 16-10 regimes (palpation: 2-3%, radiology: 27-65%). Vertebra number 26 (located beneath the dorsal fin) was the most frequently affected vertebra in smolts, while vertebra number 43 (located above the anal fin) was most frequently affected in adults.

Growth and osmoregulation in Atlantic salmon (Salmo salar ) smolts in response to different feeding frequencies and salinities

Groups of Atlantic salmon (Salmo salar) yearling smolts were reared in duplicate tanks supplied with freshwater or seawater, and subjected to different feeding frequencies, 100% (fed every day), 50% (fed every other day), 25% (fed every forth day) and 0% (starved), from 26 May to 26 July. After 8 weeks, all the groups were re-fed in excess for 6 weeks. Fish were maintained on their respective a priori salinity treatments during the 6-week follow-up period. Starvation for a period of 8 weeks in freshwater resulted in a loss of hypo-osmoregulatory ability when smolts were challenged with seawater and unfed smolts maintained in freshwater were unable to adapt to seawater in mid-July. Ration levels influenced the growth rate and body size significantly. The overall growth rate was higher in freshwater than at corresponding rations in seawater. Partial compensatory growth was observed in the 0 and 25% groups following re-feeding. Branchial Na+,K+-ATPase (NKA) activity decreased rapidly in unfed smolts in freshwater and was the lowest in the starved group, whereas an initial increase was observed in those groups reared in seawater. After re-feeding NKA activity differences decreased between the former feeding groups. Our results suggest that nutritional factors and/or energy levels are critical for the maintenance of hydro-mineral balance of salmon smolts.

Swimming suppresses hepatic vitellogenesis in European female silver eels as shown by expression of the estrogen receptor 1, vitellogenin1 and vitellogenin2 in the liver

BackgroundWhen European silver eels (Anguilla anguilla) venture into the Atlantic Ocean for their 6,000 km semelparous spawning run to the Sargasso Sea, they are still in a prepubertal stage. Further sexual development appears to be blocked by dopaminergic inhibition of hypothalamus and pituitary activity. Recently, we found that swimming for several weeks in freshwater stimulated the incorporation of fat droplets in the oocytes. So, it was hypothesized that long term swimming in seawater would release the inhibition further and would also stimulate the production of vitellogenin by the liver.MethodsFor this study a swim-flume was constructed to allow simulated migration of migratory female silver eels for 3 months (1,420 km) in natural seawater at 20 degrees C. Primers were designed for polymerase chain reactions to measure the mRNA expression of estrogen receptor 1 (esr1), vitellogenin1 (vtg1) and vitellogenin2 (vtg2) genes in the liver of European female silver eels.ResultsIn comparison to resting eels, swimming eels showed a diminished expression of esr1, vtg1 and vtg2 in the liver. They also had lower plasma calcium (Ca; indicative of vitellogenin) levels in their blood. This showed that vitellogenesis is more strongly suppressed in swimming than in resting eels. However, when eels were subsequently stimulated by 3 weekly carp pituitary extract injections, the expression of the same genes and plasma levels of Ca strongly increased in both groups to similar levels, thus equalizing the initial differences between resting and swimming.ConclusionsIt is concluded that vitellogenesis remains suppressed during resting and even more during swimming. The fact that swimming stimulates fat deposition in the oocytes but suppresses vitellogenesis indicates that these events are separated in nature and occur sequentially. Swimming-suppressed vitellogenesis may imply that in nature eels undergo vitellogenesis and final maturation near or at the spawning grounds.

The impact of different water gas levels on cataract formation, muscle and lens free amino acids, and lens antioxidant enzymes and heat shock protein mRNA abundance in smolting Atlantic salmon, Salmo salar L.

The present experiment was conducted to examine if freshwater (FW) oxygen and carbon dioxide regimes cause physiological responses that lead to cataract formation in Atlantic salmon ( Salmo salar L.) smolt. Duplicate groups of 50 g Atlantic salmon smolts were exposed to three freshwater oxygen saturation regimes (95, 112 or 125% saturation), with or without addition of carbon dioxide (measured 17–18 and 2–3 mg L − 1 , respectively), for six weeks before transfer to seawater (SW). The FW exposure groups were followed up for another six weeks under a common SW regime. Fish were screened for cataract and sampled accordingly, at start, after 6 weeks in FW and after 6 weeks in SW. Increased growth related cataract incidences and severities were recorded in SW, mainly in the groups previously exposed to normoxic and hyperoxic conditions in FW, as compared to the respective groups added carbon dioxide. The concentration of histidine compounds (imidazoles) in muscle and lens tissue, used as quantitative risk markers of cataract, were lower than observed in earlier studies, however, neither were affected by the present water gas regimes in FW nor after follow up in SW. Independently of water oxygenation in FW, muscle free amino acid profiles in salmon groups concomitantly exposed to elevated carbon dioxide indicated use of selected free amino acids for energy purposes. Significantly lower abundance of heat shock protein 70 mRNA and trends towards stepwise reduction of antioxidant enzymes mRNA in the lens from fish exposed to increased water oxygenation were recorded, probably linked to increased growth and/or external stress during smoltification. This represents a first communication on using early molecular markers to express reduced protection of the fish lens against external stress to explain cataract development.

Larval salinity tolerance of the South American salt-marsh crab, Neohelice (Chasmagnathus) granulata: physiological constraints to estuarine retention, export and reimmigration

The semiterrestrial crab Neohelice (=Chasmagnathus) granulata (Dana 1851) is a predominant species in brackish salt marshes, mangroves and estuaries. Its larvae are exported towards coastal marine waters. In order to estimate the limits of salinity tolerance constraining larval retention in estuarine habitats, we exposed in laboratory experiments freshly hatched zoeae to six different salinities (5–32‰). At 5‰, the larvae survived for a maximum of 2 weeks, reaching only exceptionally the second zoeal stage, while 38% survived to the megalopa stage at 10‰. Shortest development and negligible mortality occurred at all higher salt concentrations. These observations show that the larvae of N. granulata can tolerate a retention in the mesohaline reaches of estuaries, with a lower limit of ca. 10–15‰. Maximum survival at 25‰ suggests that polyhaline conditions rather than an export to oceanic waters are optimal for successful larval development of this species. In another experiment, we tested the capability of the last zoeal stage (IV) for reimmigration from coastal marine into brackish waters. Stepwise reductions of salinity during this stage allowed for moulting to the megalopa at 4–10‰. Although survival was at these conditions reduced and development delayed, these results suggest that already the zoea-IV stage is able to initiate the reimmigration into estuaries. After further salinity reduction, megalopae survived in this experiment for up to >3 weeks in freshwater, without moulting to juvenile crabs. In a similar experiment starting from the megalopa stage, successful metamorphosis occurred at 4–10‰, and juvenile growth continued in freshwater. Although these juvenile crabs showed significantly enhanced mortality and smaller carapace width compared to a seawater control, our results show that the late larval and early juvenile stages of N. granulata are well adapted for successful recruitment in brackish and even limnetic habitats.

Influence of dietary l-selenomethionine exposure on the survival and osmoregulatory capacity of white sturgeon in fresh and brackish water

Near simultaneous exposure to selenium and salinity stress is an environmental reality faced by white sturgeon juveniles in the San Francisco Bay-Delta, having the potential for adverse synergistic impacts on juveniles during seaward migration. White sturgeon juveniles (36–132 g) were transferred into higher salinities (15–20 ppth) after exposure to dietary selenium (0.4, 9.6, 20.5, 41.7, 89.8, or 191.1 μg Se/g diet) in the form of l-selenomethionine (SeMet) for 8 weeks in freshwater (0 ppth). White sturgeon fed above 41.7 μg Se/g diet experienced significant (P < 0.05) declines in survival relative to sturgeon fed between 0.4 and 41.7 μg Se/g diet when transferred into brackish water (15 ppth salinity) for 3 days. Plasma osmolality and ionic concentrations in white sturgeon after abrupt transfer correlated positively with dietary Se concentrations. Transfer of white sturgeon fed above 41.7 μg Se/g diet into 20 ppth brackish water for 1 day resulted in significant declines in survival relative to white sturgeon fed 0.4–41.7 μg Se/g diet. Hematocrit and dehydrational weight loss were significantly correlated with dietary Se concentrations while plasma osmolality and ion concentrations were not. The transfer of sturgeon into 20 ppth water for 3 days resulted in severe survival declines in all treatment groups. The results of this study suggest that the previously established threshold for Se toxicity (10–20 μg Se/g) is sufficient to protect white sturgeon from the adverse interactive affects resulting from increases in dietary Se and ambient water salinity.

A multivariate study on the effects of dietary vitamin K, vitamin D3 and calcium, and dissolved carbon dioxide on growth, bone minerals, vitamin status and health performance in smolting Atlantic salmon Salmo salar L.

AbstractBone deformities represent an increasing, unsolved problem for intensive farming of Atlantic salmon, Salmo salar L. To increase insight into nutritional and environmental impacts on bone mineralization, 16 groups of Atlantic salmon parr were fed diets with different levels of menadione sodium bisulphite (MSB), vitamin D3, and calcium (Ca), and exposed to different levels of dissolved carbon dioxide (CO2) based on a multivariate factorial design (24). The composition of the experimental diets was either a high or low level of each of the design variables. In addition, four groups (centre points) were fed a diet with intermediate levels of all dietary parameters and exposed to an intermediate level of dissolved CO2 to record the variation among tanks. After 6 weeks in fresh water, fish were marked according to group, transferred to a common net pen in sea water and fed commercial feed for another 11 weeks. High levels of CO2 decreased fish growth both in fresh water and sea water. No visible bone deformities were recorded at any time. Dry matter, protein and ash compositions of bone and concentrations of Ca, phosphorus (P) and magnesium (Mg) in bone were not significantly affected by any of the design variables. Minor effects were observed in clinical parameters recording health performance, stress and plasma Ca homeostasis. The mean level of vitamin D3 measured as cholecalciferol in fish organs increased significantly during the freshwater period and reflected dietary vitamin D3, whereas the level decreased when fish were fed commercial feed in sea water. The level of menaquinone in liver increased significantly in response to high dietary levels of MSB, indicating that dietary MSB was converted to menaquinone in salmon.

The effect of dietary sodium chloride on some osmoregulatory parameters of the teleost, Oreochromis niloticus, after transfer from freshwater to seawater.

The aim of this work was to determine the effects of supplemental dietary sodium chloride on salt water acclimation of tilapia Oreochromis niloticus. Fish were fed a basal diet supplemented with NaCl (8%) during three weeks in fresh water (FW) and then transferred to salt water (SW) at 15 and 20‰. Changes in plasma osmolality, chloride ion concentration (Cl−), plasma level of cortisol and gill Na+, K+-ATPase activity were measured at 6, 12, 24, 48, 72 and 168 h after transfer to 15‰SW, while the higher strength SW group (20‰) was only monitored up to 24 h. Morphological changes in the gill mitochondria-rich (MR) cells were examined in relation to environmental salinity. The changes associated with dietary NaCl were sporadic and of small magnitude. The plasma osmolality and Cl− increased immediately after transfer up to 12–24 h, but fish fed dietary salt (S) showed lower values than the control group (C). The S group showed higher plasma levels of cortisol than the control, which maintained its initial levels during the experiment. Gill Na+, K+-ATPase activity of the S group began to increase in the first hours after transfer, reaching maximum at 12 h and returned to basal level at 24 h, while the control group maintained basal levels. The differences between gill Na+, K+-ATPase activity of S and C fish were significant (p < 0.05) at 12 h. Transmission electron microscopy (TEM) revealed that MR cells in SW show more mitochondria and a more developed tubular system arising from the basolateral membrane. The MR cells of both groups frequently formed a multicellular complex in SW, consisting of a main MR and one or more accessory cells. Such complexes are rarely observed in FW. Some MR cells of fish fed supplemented dietary salt displayed convex apical membrane in FW.

The influence of sub-lethal levels of zinc on smoltifying Atlantic salmon Salmo salar and on their subsequent susceptibility to infection with Lepeophtheirus salmonis

Smoltifying Atlantic salmon were treated for 6 weeks in freshwater with 0, 200, and 400 ppb zinc (as Zn SO4). After 6 weeks salmon were transferred to a salt water seapen and exposed to infection with Lepeophtheirus salmonis for 14 weeks. Zinc treatment resulted in some physiological changes consistent with increased stress, such as decreased leukocrit values, and increased plasma cortisol levels. Plasma glucose levels were significantly elevated in fish previously treated with zinc, but this was only evident after fish had been in a seapen, 14 weeks subsequent to treatment. Improved conversion of tetraiodothyronine to triiodothyronine, as well as no changes in interrenal cell nuclear diameters suggested that overall stress effects were low. Fish exposed to 200 ppb and 400 ppb zinc showed gill pathology. Infection intensity with L. salmonis was significantly higher on salmon previously exposed to 400 ppb zinc. This research suggests that for smoltifying Atlantic salmon, the No Observed Pathological Effect (NOPE) level for exposure for ZNSO 4 may be around 150 ppb.

Effects of a dietary betaine/amino acid additive on growth and seawater adaptation in yearling chinook salmon

The effect of FinnStim, a betaine/amino acid mixture, on growth and seawater adaptation was studied in yearling chinook salmon Oncorhynchus tshawytscha. The fish were marked with PIT tags and divided into two groups. One group was fed a diet with 1% FinnStim added and the other was fed a control diet with 1% α-cellulose. The diets were fed for 6 weeks in fresh water at a level of 1% body weight per day and for 8 weeks in seawater netpens at 3% body weight per day. Seawater challenge tests were conducted three times during the freshwater phase of the experiment. Growth and mortality in fresh water did not differ significantly between treatments. Seawater challenge tests at days 5, 47 and 70 of freshwater rearing revealed no significant difference in plasma sodium concentration between groups. During the seawater phase of the experiment, the group receiving FinnStim had a significantly higher growth rate and a significantly lower concentration of plasma sodium at the end of the study. The improved growth performance in the group receiving FinnStim was due to a reduction in the number of salmon growing poorly.