Freshwater Smolts Research Articles

Red and melanized focal changes in white skeletal muscle in Atlantic salmon (Salmo salar): Comparative analysis of farmed, wild and hybrid fish reared under identical conditions.

Selective breeding plays a vital role in the production of farmed Atlantic salmon and has shown success in many aspects. Still, challenges related to fish health and welfare continue to result in significant economic losses. One such challenge is red and melanized focal changes (RFC/MFC), which result from acute and chronic inflammation, respectively, in the skeletal muscle. Importantly, RFC/MFC has not been observed in wild Atlantic salmon, suggesting that both external and genetic factors may contribute to the development of inflammation. To investigate the underlying cause of RFC/MFC, we conducted a study involving 1854 Atlantic salmon of farmed, wild and hybrid origin. All fish were reared under identical conditions to minimize the influence of external factors. Throughout the production cycle, the fish was monitored for growth parameters and examined for RFC/MFC using macroscopic and histological analysis. We found no association between the experimental groups and the presence of RFC/MFC. Histological investigations revealed melano-macrophages in the soft tissue in freshwater smolt, although no macroscopic discoloration was observed. MFC showed granulomas in various stages, suggesting a complex progression of the condition. In summary, we conclude that RFC/MFC is primarily caused by external factors found in the rearing facilities of farmed Atlantic salmon.

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.

Tailoring freshwater diets towards boosted immunity and pancreas disease infection robustness in Atlantic salmon post smolts

The aim of the current study was to investigate how freshwater diets impact on immunity in Atlantic salmon smolts in freshwater, during transfer to seawater and in post smolts during the seawater stage with and without pancreas disease (PD) infection. Three specific freshwater diets were prepared: (i) A diet similar in composition to commercial salmon freshwater diets (Standard diet); (ii) A diet composed of vegetable oils (rapeseed, palm and linseed oils) mimicking the fat composition in aquatic insects – the natural diet of wild salmon in freshwater (Fatty acid diet); (iii) A diet enriched with possible immune modulating amino acids including dl-methionine, l-lysine, l-threonine and taurine (Amino acid diet). After seawater transfer, all fish were fed the same commercial diet. Head kidneys were extracted, and their leukocytes isolated from smolts right before transfer to seawater, from post smolts one and six weeks after transfer to seawater, and from post smolts in seawater after 8 weeks of ongoing PD infection. In addition, to provoke bacterial or virus induced inflammation in vitro, the individual leukocyte suspension from all fish were stimulated by lipopolysaccharide (LPS) or polyinosinic acid: polycytidylic acid (PIC).The transfer of smolts from fresh-to seawater changed the transcription of several types of genes. Particularly in isolates from fish fed the Standard or Fatty acid diet in freshwater, overall gene transcription (IL-1β, CD83, INF-γ, cox2, cd36, MGAT2, catalase) declined. However, the Amino acid diet stimulated the LPS induced gene transcription of IL-1β, CD83, Cox2, and INF-γ at this stage. In freshwater smolts, PIC stimulated leukocytes showed higher transcription level of Mx and viperin in the Fatty acid and Amino acid diet groups compared to the Standard diet group. In seawater post smolts, Mx and viperin responded similarly to PIC challenge in all diet groups. Furthermore, leukocytes isolated from PD infected fish, continued responding to PIC, regardless of freshwater diet.

Sulfate homeostasis in Atlantic salmon is associated with differential regulation of salmonid-specific paralogs in gill and kidney.

Sulfate (SO42‐) regulation is challenging for euryhaline species as they deal with large fluctuations of SO42‐ during migratory transitions between freshwater (FW) and seawater (SW), while maintaining a stable plasma SO42‐ concentration. Here, we investigated the regulation and potential role of sulfate transporters in Atlantic salmon during the preparative switch from SO42‐ uptake to secretion. A preparatory increase in kidney and gill sodium/potassium ATPase (Nka) enzyme activity during smolt development indicate preparative osmoregulatory changes. In contrast to gill Nka activity a transient decrease in kidney Nka after direct SW exposure was observed and may be a result of reduced glomerular filtration rates and tubular flow through the kidney. In silico analyses revealed that Atlantic salmon genome comprises a single slc13a1 gene and additional salmonid‐specific duplications of slc26a1 and slc26a6a, leading to new paralogs, namely the slc26a1a and ‐b, and slc26a6a1 and ‐a2. A kidney‐specific increase in slc26a6a1 and slc26a1a during smoltification and SW transfer, suggests an important role of these sulfate transporters in the regulatory shift from absorption to secretion in the kidney. Plasma SO42‐ in FW smolts was 0.70 mM, followed by a transient increase to 1.14 ± 0.33 mM 2 days post‐SW transfer, further decreasing to 0.69 ± 0.041 mM after 1 month in SW. Our findings support the vital role of the kidney in SO42‐ excretion through the upregulated slc26a6a1, the most likely secretory transport candidate in fish, which together with the slc26a1a transporter likely removes excess SO42‐, and ultimately enable the regulation of normal plasma SO42‐ levels in SW.

Effects of elevated temperature on osmoregulation and stress responses in Atlantic salmon Salmo salar smolts in fresh water and seawater.

Smolting in Atlantic salmon Salmo salar is a critical life-history stage that is preparatory for downstream migration and entry to seawater that is regulated by abiotic variables including photoperiod and temperature. The present study was undertaken to determine the interaction of temperature and salinity on salinity tolerance, gill osmoregulatory proteins and cellular and endocrine stress in S. salar smolts. Fish were exposed to rapid changes in temperature (from 14 to 17, 20 and 24°C) in fresh water (FW) and seawater (SW), with and without prior acclimation and sampled after 2 and 8 days. Fish exposed simultaneously to SW and 24°C experienced 100% mortality, whereas no mortality occurred in any of the other groups. The highest temperature also resulted in poor ion regulation in SW with or without prior SW acclimation, whereas no substantial effect was observed in FW. Gill Na+ -K+ -ATPase (NKA) activity increased in SW fish compared to FW fish and decreased with high temperature in both FW and SW. Gill Nkaα1a abundance was high in FW and Nkaα1b and Na+ -K+ -2Cl- cotransporter high in SW, but all three were lower at the highest temperature. Gill Hsp70 levels were elevated in FW and SW at the highest temperature and increased with increasing temperature 2 days following direct transfer to SW. Plasma cortisol levels were elevated in SW at the highest temperature. Our results indicate that there is an important interaction of salinity and elevated temperature on osmoregulatory performance and the cellular stress response in S. salar, with an apparent threshold for osmoregulatory failure in SW above 20°C.

Atlantic salmon skin barrier functions gradually enhance after seawater transfer

Atlantic salmon farming operates with high production intensities where skin integrity is recognized as a central factor and indicator for animal health and welfare. In the described trial, the skin development and its immune status in healthy Atlantic salmon reared in two different systems, a traditional open net-pen system and a semi-closed containment system, were investigated. Freshwater smolts were compared to post-smolts after 1 and 4 months in seawater. Growth performance, when adjusted for temperature, was equal between the systems. Skin analyses, including epidermis and dermis, showed that thickness and mucus cell numbers increased in pace with the growth and time post seawater transfer (PST). Gene expression changes suggested similar processes with development of connective tissue, formation of extracellular matrix and augmented cutaneous secretion, changes in mucus protein composition and overall increased immune activity related to gradually enforced protection against pathogens. Results suggest a gradual morphological development in skin with a delayed recovery of immune functions PST. It is possible that Atlantic salmon could experience increased susceptibility to infectious agents and risk of diseases during the first post-smolt period.

Assessment of interbreeding and introgression of farm genes into a small Scottish Atlantic salmon Salmo salar stock: ad hoc samples - ad hoc results?

An eclectic set of tissues and existing data, including purposely collected samples, spanning 1997-2006, was used in an ad hoc assessment of hybridization and introgression of farmed wild Atlantic salmon Salmo salar in the small Loch na Thull (LnT) catchment in north-west Scotland. The catchment is in an area of marine farm production and contains freshwater smolt rearing cages. The LnT S. salar stock was found to be genetically distinctive from stocks in neighbouring rivers and, despite regular reports of feral farm S. salar, there was no evidence of physical or genetic mixing. This cannot be completely ruled out, however, and low level mixing with other local wild stocks has been suggested. The LnT population appeared underpinned by relatively smaller effective number of breeders (Neb ) and showed relatively low levels of genetic diversity, consistent with a small effective population size. Small sample sizes, an incomplete farm baseline and the use of non-diagnostic molecular markers, constrain the power of the analysis but the findings strongly support the LnT catchment having a genetically distinct wild S. salar population little affected by interbreeding with feral farm escapes.

Initiation of migration and movement rates of Atlantic salmon smolts in fresh water

Timing of ocean entry is critical for marine survival of both hatchery and wild Atlantic salmon (Salmo salar) smolts. Management practices and barriers to migration such as dams may constrain timing of smolt migrations resulting in suboptimal performance at saltwater entry. We modeled influences of stocking location, smolt development, and environmental conditions on (i) initiation of migration by hatchery-reared smolts and (ii) movement rate of hatchery- and wild-reared Atlantic salmon smolts in the Penobscot River, Maine, USA, from 2005 through 2014 using acoustic telemetry data. We also compared movement rates in free-flowing reaches with rates in reaches with hydropower dams and head ponds. We compared movement rates before and after (1) removal of two mainstem dams and (2) construction of new powerhouses. Initiation of movement by hatchery fish was influenced by smolt development, stocking location, and environmental conditions. Smolts with the greatest gill Na+, K+-ATPase (NKA) activity initiated migration 24 h sooner than fish with the lowest gill NKA activity. Fish with the greatest cumulative thermal experience initiated migration 5 days earlier than those with lowest cumulative thermal experience. Smolts released furthest from the ocean initiated migration earlier than those released downstream, but movement rate increased by fivefold closer to the ocean, indicating behavioral trade-offs between initiation and movement rate. Dams had a strong effect on movement rate. Movement rate increased from 2.8 to 5.4 km·h−1in reaches where dams were removed, but decreased from 2.1 to 0.1 km·h−1in reaches where new powerhouses were constructed. Movement rate varied throughout the migratory period and was inversely related to temperature. Fish moved slower at extreme high or low discharge. Responses in fish movement rates to dam removal indicate the potential scope of recovery for these activities.

Plasma growth hormone-binding protein levels in Atlantic salmon Salmo salar during smoltification and seawater transfer.

Specific growth hormone (GH)-binding protein (Ghbp) was purified from Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss plasma with immunoprecipitation and characterized in cross-linking studies using autoradiography and western blots. The size of the Ghbp was estimated to be c. 53 kDa. A radioimmunoassay was established to measure Ghbp in salmonids, using antibodies specific against the extracellular segment of the S. salar growth hormone receptor 1 (grh1; GenBank AY462105). Plasma Ghbp levels were measured in S. salar smolts in fresh water and after transfer to seawater (SW; experiments 1 and 2), and in post-smolts kept at different salinities (0, 12, 22 and 34) for 3 months (experiment 3). A transient increase in plasma Ghbp, which lasted for 1 month or less, was noted in smolts after transfer to SW. Concomitantly, plasma GH and gill Na(+) -K(+) -ATPase activity increased during smoltification (in experiment 2). No difference in plasma Ghbp was evident between post-smolts kept at different salinities, although the fish kept at salinity 34 had higher plasma GH than the group kept at salinity 22 and higher hepatic ghr1 expression than post-smolts kept at salinity 12. This suggests that plasma Ghbp regulation may respond to salinity changes in the short term. The lack of correlation between Ghbp, plasma GH and hepatic ghr1 expression in the long-term post-smolt experiment indicates that Ghbp levels may be regulated independently of other components of the endocrine GH system in salmonids.

Atlantic salmon (Salmo salar L.) smolts require more than two weeks to recover from acidic water and aluminium exposure

The detrimental effects of acid rain and aluminium (Al) on salmonids have been extensively studied, yet knowledge about the extent and rate of potential recovery after exposures to acid and Al episodes is limited. Atlantic salmon smolts in freshwater (FW) were exposed for 2 and 7-day episodes (ACID2 and ACID7, respectively) to low pH (5.7±0.2) and inorganic aluminium (Ali; 40±4μg) and then transferred to good water quality, control water (CW; pH 6.8±0.1; <14±2μg Ali). Al accumulation on gills after 2 and 7 days of acid/Al exposure was 35.3±14.1 and 26.6±1.8μgg−1 dry weight, respectively. These elevated levels decreased 2 days post transfer to CW and remained higher than in control (CON; 5–10μg Ali) for two weeks. Plasma Na+ levels in ACID2 and ACID7 smolts decreased to 141±0.8 and 138.6±1.4mM, respectively, and remained significantly lower than CON levels for two weeks post transfer to CW. Similarly, plasma Cl− levels in ACID7 smolts (124.3±2.8mM) were significantly lower than in CON, with Cl− levels remaining significantly lower in ACID7 (126.2±4.8mM) and ACID2 (127.6±3.7mM) than in CON following 9 and 14 days post-transfer to CW, respectively. ACID2 and ACID7 smolts sustained elevated plasma glucose levels post transfer to CW suggesting elevated stress for more than a week following exposure. While gill Na+, K+-ATPase (NKA) activity was only slightly affected in ACID2 and not in ACID7 smolts in FW, acid/Al exposure resulted in a transient decrease in NKA activity following SW exposure in both groups. Acid/Al episodes had limited impact on isoform specific NKA α-subunit mRNA during exposure. However, the transfer of ACID2 and ACID7 smolts to CW showed an increase in NKAα1a mRNA (the FW isoform) and inhibited the up-regulation of NKAα1b (the SW isoform), probably resulting in higher abundance of the enzyme favouring ion uptake. Gill caspase 3B gene transcription did not change in acid/Al treated smolts, indicating no increased apoptosis in gills. ACID2 and ACID7 treatments resulted in lower smolt-related gill transcription of the gene encoding the tight junction protein claudin 10e compared to CON, while the gene encoding claudin 30 showed lower mRNA expression only after 11 days SW exposure in ACID7 fish. Our data suggest that acid/Al conditions affect ion perturbations through a combination of alteration of the preparatory increase in paracellular permeability and negative impact on the SW type NKA α-subunit mRNA transcripts, and raise major concerns regarding the recovery of physiological disruption in smolts following acid/Al exposure. Smolts may require more than two weeks to fully recover from even short moderate episodes of acid/Al exposure. Acid/Al exposure thus probably has greater impact on salmon populations than previously acknowledged.

Estrogenic compounds decrease growth hormone receptor abundance and alter osmoregulation in Atlantic salmon

Exposure of Atlantic salmon smolts to estrogenic compounds is shown to compromise several aspects of smolt development. We sought to determine the underlying endocrine mechanisms of estrogen impacts on the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis. Smolts in freshwater (FW) were either injected 3 times over 10days with 2μgg−1 17β-estradiol (E2) or 150μgg−1 4-nonylphenol (NP). Seawater (SW)-acclimated fish received intraperitoneal implants of 30μgg−1 E2 over two weeks. Treatment with these estrogenic compounds increased hepatosomatic index and total plasma calcium. E2 and NP reduced maximum growth hormone binding by 30–60% in hepatic and branchial membranes in FW and SW, but did not alter the dissociation constant. E2 and NP treatment decreased plasma levels of IGF-I levels in both FW and SW. In FW E2 and NP decreased plasma GH whereas in SW plasma GH increased after E2 treatment. Compared to controls, plasma chloride concentrations of E2-treated fish were decreased 5.5mM in FW and increased 10.5mM in SW. There was no effect of NP or E2 on gill sodium–potassium adenosine triphosphatase (Na+/K+-ATPase) activity in FW smolts, whereas E2 treatment in SW reduced gill Na+/K+-ATPase activity and altered the number and size of ionocytes. Our data indicate that E2 downregulates the GH/IGF-I-axis and SW tolerance which may be part of its normal function for reproduction and movement into FW. We conclude that the mechanism of endocrine disruption of smolt development by NP is in part through alteration of the GH/IGF-I axis via reduced GH receptor abundance.

PCD Risk 1: A tool for assessing and reducing ecological risks of hatchery operations in freshwater

PCD RISK 1 is a flexible computer model designed to assist in assessment and reduction of ecological risks to natural origin fish from hatchery fish releases. PCD RISK 1 simulates predation, competition, and disease impacts on naturally produced salmonids caused by hatchery smolts in fresh water as they move downstream or residualize after release. Individual- based, the model relies upon user specified inputs of up to 45 variables, such as number and size of hatchery and wild fish, and water temperature. The model generates hatchery and natural fish of specified size distributions, then randomly pairs them for interactions for a specified number of days and encounters. Wild fish are subjected to predation if they are less than 50% the length of hatchery fish, otherwise they are subjected to competition. After all of the allowable competition and predation occurs, survivors are subjected to disease risk. The model can provide either deterministic or probabilistic output. Deterministic output includes the number and proportion of wild fish that die from predation, competition, disease, and from all interactions combined. Probabilistic output includes probability distributions of the number and proportion of mortalities, based on user-specified uncertainty input either as uniform or triangular distributions for any of several variables. If desired, the model can also quantify impacts for multiple age or size classes. Simulations of a variety of hatchery programs reveal that risks are highly diverse and that a variety of options are available to reduce risks.

Gill Na+,K+-ATPase of Atlantic salmon smolts in freshwater is not a predictor of long-term growth in seawater

Gill Na+,K+-ATPase activity is a widely used measure of osmoregulatory preparedness in salmonid smolts. The degree to which this measure may predict long term performance is uncertain. In order to assess the relationship of this enzyme to long term growth and ion homeostasis, a cohort of Atlantic salmon hatchery smolts was used in a controlled environment with no salinity perturbations. In May 2006, gill Na+,K+-ATPase activity from 940 individually PIT tagged, Penobscot River smolts (USFWS, Green Lake National Fish Hatchery, Maine, United States) was measured immediately prior to isothermal transfer from freshwater to 32ppt seawater. From the observed range of activities, individuals were classified as having “low”, “middle”, or “high” enzyme activity levels. Individual size (fork length and mass) was recorded on days 0, 1, 3, and 14 and monthly for four months. Growth rates over four time periods were calculated for individual fish maintained until the end of the experiment. Gill Na+,K+-ATPase activities were also measured from a subset of sampled fish. All groups effectively osmoregulated as evidenced by minor perturbations in plasma osmolyte levels. Apart from initial weight loss on transfer, fish grew throughout the experiment, however, there were no differences (fish size, growth rate, and gill Na+,K+-ATPase activity in seawater) among groups with initially different gill Na+,K+-ATPase activities (prior to seawater entry). While gill Na+,K+-ATPase activity may be predictive of performance during the acute phase of acclimation (first few days), typical variation in this enzyme, expressed in freshwater at the peak of smolting, does not appear to be predictive of long-term growth in seawater.

Effects of environmental salinity, biopsy, and GH and IGF-I administration on the expression of immune and osmoregulatory genes in the gills of Atlantic salmon (Salmo salar)

Immune-endocrine interactions are thought to be important for fish health, especially following injury of peripheral tissues. We investigated the influences of gill biopsy (surgical removal of several gill filaments with minimal bleeding) on mRNA levels of osmoregulatory and immune genes in peripheral tissues of Atlantic salmon (Salmo salar) smolts in freshwater (FW) and seawater (SW). The effects of growth hormone (GH) and insulin-like growth factor (IGF)-I, which are known as both osmoregulatory and immunomodulatory hormones, were also examined in FW- and SW-acclimated fish. Compared with the intact gill arch of the same fish, the residuary filaments after biopsy showed significant reduction in Na+,K+-ATPase (NKA) and cystic fibrosis transmembrane conductance regulator (CFTR) mRNA levels in FW fish. In contrast, mRNA levels of these genes in the filaments with biopsy were increased in SW fish. Increased caspase gene transcription was observed in the cut filaments of SW fish, but not in those of FW fish. Lysozyme gene transcription was stimulated in the residuary gill filaments after biopsy in both FW and SW. Administration of salmon GH or IGF-I increased mRNA levels of NKA, CFTR, caspase, and lysozyme in the gills after biopsy in FW fish. In SW-acclimated fish, there was no significant effect of GH or IGF-I on the transcription of osmoregulatory or immune genes. The results in this study showed tight relationships between immunity, osmoregulation, and endocrine system in fish gills, and that nonlethal gill biopsy can be used for investigations of fish health.

DIURNAL EXPRESSION OF CLOCK GENES IN PINEAL GLAND AND BRAIN AND PLASMA LEVELS OF MELATONIN AND CORTISOL IN ATLANTIC SALMON PARR AND SMOLTS

In Atlantic salmon, the preadaptation to a marine life, i.e., parr-smolt transformation, and melatonin production in the pineal gland are regulated by the photoperiod. However, the clock genes have never been studied in the pineal gland of this species. The aim of the present study was to describe the diurnal expression of clock genes (Per1-like, Cry2, and Clock) in the pineal gland and brain of Atlantic salmon parr and smolts in freshwater, as well as plasma levels of melatonin and cortisol. By employing an out-of-season smolt production model, the parr-smolt transformation was induced by subjecting triplicate groups of parr to 6 wks (wks 0 to 6) under a 12 h:12 h light-dark (LD) regime followed by 6 wks (wks 6 to 12) of continuous light (LL). The measured clock genes in both pineal gland and brain and the plasma levels of melatonin and cortisol showed significant daily variations in parr under LD in wk 6, whereas these rhythms were abolished in smolts under LL in wk 12. In parr, the pineal Per1-like and Cry2 expression peaked in the dark phase, whereas the pineal Clock expression was elevated during the light phase. Although this study presents novel findings on the clock gene system in the teleost pineal gland, the role of this system in the regulation of smoltification needs to be studied in more detail. (Author correspondence: ti.hu@hotmail.com)

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.

Effects of hexazinone and atrazine on the physiology and endocrinology of smolt development in Atlantic salmon

Exposure to hexazinone (HEX) and atrazine (ATZ), highly mobile and widely used herbicides along rivers in the United States, is potentially harmful to Atlantic salmon, which have been listed as an endangered species. To determine the effects of these contaminants on smolt development, juvenile Atlantic salmon were exposed under flow-through conditions to 100 μg l −1 HEX, 10 and 100 μg l −1 ATZ in fresh water (FW) for 21 days at 10 °C beginning in mid-April. Twelve fish per treatment were sampled in FW, following a 24 h seawater (SW) challenge and after growth for 3 months in SW. Exposure to 100 μg l −1 HEX or 10 μg l −1 ATZ caused no mortalities of smolts in FW or after SW challenge, while 9% of the fish exposed to 100 μg l −1 ATZ died during exposure. Fish exposed to 100 μg l −1 ATZ reduced feeding after 10 days of exposure and had an impaired growth rate in FW and during the first month in SW; compensatory growth occurred in the second and third month in SW. HEX and ATZ at 10 μg l −1 exposure had no effect on plasma levels of cortisol, growth hormone (GH), insulin growth factor I (IGF-I), thyroxine (T 4) and plasma 3,5,3′-triiodo- l-thyronine (T 3), Cl −, Mg 2+, Na +, Ca 2+ in FW or after SW challenge. FW smolts exposed to 100 μg l −1 ATZ had decreased plasma Cl −, Mg 2+, Na + and Ca 2+ ions and increased cortisol. No effect on plasma levels of GH, IGF-I, T 4 or T 3 was found in FW smolts exposed to 100 μg l −1 ATZ. Following SW challenge, fish previously exposed to 100 μg l −1 ATZ had significant increases in hematocrit, plasma cortisol, Cl −, Mg 2+, Na +, Ca 2+ and a decrease in T 4 and T 3. It is concluded that under the conditions imposed in this study, HEX does not affect salinity tolerance of Atlantic salmon smolts, while ATZ causes ionoregulatory, growth and endocrine disturbance.

Influence of Oxygen on the Toxicity of Chloramine‐T to Atlantic Salmon Smolts in Freshwater and Seawater

AbstractChloramine‐T is a widely used disinfectant for the treatment of gill diseases of fish in freshwater, and more recently attention has turned to its use in seawater. However, despite the wide use of chloramine‐T, few studies have examined its acute toxicity to fish. In this study we examined the acute (within 12 h) toxicity of freshwater‐ and seawater‐acclimated Atlantic salmon Salmo salar smolts under aerated (100% air saturation with O2) and oxygen supersaturation conditions (200% air saturation with O2). Chloramine‐T was more acutely toxic to salmon in seawater than to those in freshwater, and oxygen supersaturation enhanced the toxicity in both sea‐ and freshwater. In aerated freshwater the median lethal times (LT50s) for chloramine‐T concentrations of 50 and 25 mg/L were 166.8 and 474.3 min, respectively; in aerated seawater they were 119.1 and 297.3 min, respectively. However, in freshwater at 200% air saturation with oxygen, the 50 and 25 mg/L LT50s were 133.6 and 190.9 min, respectively; in seawater at 200% air oxygen saturation, the values were 85.3 and 158.9 min, respectively. Freshwater Atlantic salmon appear to be as sensitive as rainbow trout Oncorhynchus mykiss and more sensitive than channel catfish Ictalurus punctatus to chloramine‐T toxicity. Seawater‐acclimated salmon, however, appear to be more sensitive to chloramine‐T than are trout and catfish. The primary mechanism of toxicity in both seawater and freshwater appears to be extensive oxidative necrosis of the gill filament and lamellar epithelium, causing acute osmoregulatory dysfunction.

The effect of atrazine on Atlantic salmon ( Salmo salar) smolts in fresh water and after sea water transfer

Groups of Atlantic salmon smolts were exposed to low levels of the pesticide atrazine (2-chloro-4-ethylamino-6-isopropylamino- S-triazine) (0–22.7 μg l −1) in fresh water and the physiological effects of exposure were measured. Further experiments exposed salmon smolts to similar levels of atrazine in fresh water, and then exposed them to full strength sea water. Atrazine in fresh water resulted in a significant reduction in gill Na +K +ATPase activity at concentrations of 2.0, 5.0 and 10.0 μg l −1. There were few other physiological changes in the smolts except for slightly elevated plasma cortisol concentrations and monovalent ion concentrations at and above 5.0 μg l −1. However, a sea water challenge caused mortalities in smolts that had been pre-exposed to atrazine in fresh water at concentrations of 1.0, 2.0, 5.0, 10.0 and 22.7 μg l −1. Moreover, surviving fish showed signs of major physiological stress: elevated plasma cortisol, thyroxine, osmolality, and monovalent ion concentrations. However, atrazine exposure had no effect on muscle or plasma water contents. The data suggests that exposure of salmon smolts to atrazine in fresh water may compromise their physiological capabilities to survive in saline conditions.

The effects of 4-nonylphenol and atrazine on Atlantic salmon (Salmo salar L) smolts

Abstract Exposure of Atlantic salmon ( Salmo salar L.) smolts in freshwater to environmental levels of the oestrogenic chemical 4-nonylphenol (4-NP) (concentrations 5–20 μl l −1 ) during the peak migration period had no significant effect on gill Na + K + ATPase activity, plasma vitellogenin (VTG) levels or hypoosmoregulatory performance as indicated by survival in sea water. However, where smolts were exposed to mixtures of 4-NP and the pesticide atrazine at concentrations of 5.0/1.0 and 10.0/2.0 μg l −1 , respectively, there were significant differences in the gill Na + K + ATPase activity, plasma Cl − and Na + and increased mortalities when transferred to seawater. Smolts retained for a short period in an estuary with high levels of oestrogenic contamination showed no significant differences in gill Na + K + ATPase activity, plasma vitellogenin (VTG) levels or hypoosmoregulatory performance compared to fish maintained in an adjacent uncontaminated estuary. The results of the study are discussed in relation to the effects of environmental levels of oestrogenic contaminants on smoltification and migration in wild salmon.