Molecular quantification of parasitic sea louse larvae depends on species and life stage.

Sea lice cause substantial economic and environmental harm to Norway's aquaculture industry and wild salmonid populations. Rapid, accurate quantification of lice larval densities in coastal waters remains the greatest bottleneck for providing empirical data on infestation risk within wild salmon habitats and aquaculture production regions. We evaluated the capability of droplet digital PCR (ddPCR) as an absolute quantification method for the planktonic stages of two parasitic louse species, Lepeophtheirus salmonis (Krøyer) and Caligus elongatus (von Nordman). Results demonstrated linear relationships between the DNA quantity measured and the number of spiked larvae for both species and life stages. However, L. salmonis contained a significantly greater number of DNA copies than C. elongatus individuals and for C. elongatus, nauplii displayed a significantly higher number of DNA copies than copepodids. Our results suggest that ddPCR can effectively enumerate louse larvae, but interpreting ddPCR results differ between the two louse species. Obtaining larval abundance estimates from marine plankton samples will depend on the nauplii to copepodid ratio for C. elongatus, but not for L. salmonis.

Data for characterization of the optical properties of Atlantic salmon (Salmo salar) blood.

Photoplethysmography is a recent addition to physio-logging in Atlantic salmon which can be used for pulse oximetry provided that the properties for light propagation in salmon tissues are known. In this work, optical properties of three constituents of Atlantic salmon blood have been characterized using a photo spectrometer in the VIS-NIR range (450-920 nm). Furthermore, Atlantic salmon blood cell size has been measured using a Coulter counter as part of light scattering property evaluations. Results indicate that plasma contributes little to scattering and absorption for wavelengths typically used in pulse oximetry as opposed to blood cells which are highly scattering. Extinction spectra for oxygenated and deoxygenated hemoglobin indicate that Atlantic salmon hemoglobin is similar to that in humans. Pulse oximetry sensors originally intended for human applications may thus be used to estimate blood oxygenation levels for this species.

Large-effect loci mediate rapid adaptation of salmon body size after river regulation.

Understanding the potential of natural populations to adapt to altered environments is becoming increasingly relevant in evolutionary research. Currently, our understanding of adaptation to human alteration of the environment is hampered by lack of knowledge on the genetic basis of traits, lack of time series, and little or no information on changes in optimal trait values. Here, we used time series data spanning nearly a century to investigate how the body mass of Atlantic salmon (Salmo salar) adapts to river regulation. We found that the change in body mass followed the change in waterflow, both decreasing to ∼1/3 of their original values. Allele frequency changes at two loci in the regions of vgll3 and six6 predicted more than 80% of the observed body mass reduction. Modeling the adaptive dynamics revealed that the population mean lagged behind its optimum before catching up approximately six salmon generations after the initial waterflow reduction. Our results demonstrate rapid adaptation mediated by large-effect loci and provide insight into the temporal dynamics of evolutionary rescue following human disturbance.

Early-season brown trout (Salmo trutta) migrants grow and survive better at sea.

The timing of seaward migration is a key life-history trait for many anadromous fish species, with growth and survival at sea depending on a match/mismatch scenario between the timing of the sea entry and optimal conditions. Based on a 25-year study with 15,226 individually tagged brown trout (Salmo trutta) in a Norwegian river, we analysed how the within-season timing of sea migration impacted growth and survival. In both first-time and veteran migrants, marine growth was highest for early migrating individuals, large individuals, and those with a low condition factor when entering the sea. Survival was highest for individuals entering the sea early in the season. In first-time migrants, survival increased with body length. Survival also increased with the number of other smolts migrating simultaneously. As the early smolts were the most successful, it may seem strange that many smolts migrate later in the season. We suggest that late-migrating smolts may not be of a size and/or physiological state suitable for early marine conditions, and may make the best of a bad situation.

Redefining the oceanic distribution of Atlantic salmon.

Determining the mechanisms driving range-wide reductions in Atlantic salmon marine survival is hindered by an insufficient understanding of their oceanic ecology and distribution. We attached 204 pop-up satellite archival tags to post-spawned salmon when they migrated to the ocean from seven European areas and maiden North American salmon captured at sea at West Greenland. Individuals migrated further north and east than previously reported and displayed increased diving activity near oceanographic fronts, emphasizing the importance of these regions as feeding areas. The oceanic distribution differed among individuals and populations, but overlapped more between geographically proximate than distant populations. Dissimilarities in distribution likely contribute to variation in growth and survival within and among populations due to spatio-temporal differences in environmental conditions. Climate-induced changes in oceanographic conditions will alter the location of frontal areas and may have stock-specific effects on Atlantic salmon population dynamics, likely having the largest impacts on southern populations.

Factors influencing return rate and marine residence duration in sea trout populations in Central Norway.

Brown trout (Salmo trutta) display extensive plasticity in marine migratory behaviours, with marine migrations considered to be an adaptive strategy which enables sea trout to maximize growth and reproductive potential. However, marine migrations are not without associated costs, including threats posed by ever-increasing salmon lice (Lepeophtheirus salmonis) infestations. In the present study, we used passive integrated transponder technology to characterize variability in sea trout migration behaviour amongst three catchments situated in a region of intensive salmon farming in central Norway. Specifically, we investigate how lice infestation, out-migration date and body size alter sea trout return rate and marine residence duration during the first out-migration to sea from each catchment. Distinct catchment-specific differences in sea trout out-migration size and the number of cohorts were observed, but larger body size did not guarantee the successful return of migrating trout. The marine residence duration of individuals that successfully returned to freshwater was positively correlated with lice infestation risk, suggesting for these individuals the lethal infestation threshold had not been reached. Our results also suggest that sea trout populations from lotic-dominated catchments are potentially at greater risk from size-related threats to their survival encountered during their marine migrations than sea trout from lentic-dominated catchments. The variability in sea trout migratory behaviour amongst catchments observed here emphasizes the challenges fisheries managers face when deciding the best actions to take to protect the anadromous portion of brown trout populations.

Repeatable individual variation in migration timing in two anadromous salmonids and ecological consequences.

Consistent individual differences in behavior have been demonstrated for many animals, but there are few studies of consequences of such repeated behavior in the wild. We tested consistency in migration timing to and from the sea among anadromous Arctic char (Salvelinus alpinus) and brown trout (Salmo trutta), using data from a study period of about 25 years, including more than 27,000 uniquely Carlin-tagged individuals that migrated to sea for feeding in the spring and returned to the river in late summer for up to 13 successive years. Consistency was found between individuals across time in timing of the seaward migration. Individuals migrating early during their first migration tended to migrate early the following years, and late migrants tended to migrate late. The same pattern was found also at ascent to freshwater. Hence, this study demonstrated that individual fish in nature can differ in behavior related to migration timing and that these differences can be consistent during their lifetime. Early migrants increased their mass more than late migrants and had a higher specific growth rate. Early migrating Arctic char, but not brown trout, experienced a longer life after the first migration to sea than late migrants. In both species, maturity occurred earlier in individuals that migrated early. For brown trout, but not for Arctic char, fecundity was significantly correlated to the timing of smolt migration. Hence, the repeatable individual variation in migration timing seemed to have ecological and fitness consequences in terms of growth, longevity, timing of maturity, and lifetime fecundity.

Effects of environmental enrichment on forebrain neural plasticity and survival success of stocked Atlantic salmon.

Fish reared for stocking programmes are severely stimulus deprived compared with their wild conspecifics raised under natural conditions. This leads to reduced behavioural plasticity and low post-release survival of stocked fish. Environmental enrichment can have positive effects on important life skills, such as predator avoidance and foraging behaviour, but the neural mechanisms underpinning these behavioural changes are still largely unknown. In this study, juvenile Atlantic salmon (Salmo salar) were reared in an enriched hatchery environment for 7 weeks, after which neurobiological characteristics and post-release survival were compared with those of fish reared under normal hatchery conditions. Using in situ hybridization and qPCR, we quantified the expression of brain-derived neurotrophic factor (bdnf) and the neural activity marker cfos in telencephalic subregions associated with relational memory, emotional learning and stress reactivity. Aside from lower expression of bdnf in the Dlv (a region associated with relational memory) of enriched salmon, we observed no other significant effects of enrichment in the studied regions. Exposure to an enriched environment increased post-release survival during a 5 month residence in a natural river by 51%. Thus, we demonstrate that environmental enrichment can improve stocking success of Atlantic salmon parr and that environmental enrichment is associated with changes in bdnf expression in the fish's hippocampus-equivalent structure.

The Phe362Tyr mutation conveying resistance to organophosphates occurs in high frequencies in salmon lice collected from wild salmon and trout.

The parasitic salmon louse, and its resistance to chemical delousing agents, represents one of the largest challenges to the salmon aquaculture industry. We genotyped lice sampled from wild salmon and sea trout throughout Norway with the recently identified Phe362Tyr mutation that conveys resistance to organophosphates. These results were compared to data from lice sampled on farmed salmon in the same regions. The resistant (R) allele was observed in salmon lice from wild salmon and sea trout throughout Norway, although its frequency was highest in farming-intense regions. In most regions, the frequency of the R allele was higher in lice collected from wild sea trout than wild Atlantic salmon, and in all regions, the frequency of the R allele was similar in lice collected from wild sea trout and farmed Atlantic salmon. The R allele is only selected for in fish-farms where organophosphates are used for delousing. Therefore, our results suggest extensive exchange of lice between farmed and wild hosts, and indicate that in farming-dense regions in Norway, aquaculture represents a major driver of salmon louse population structure. Finally, these data suggest that the wild hosts within the regions studied will not delay the spread of resistance when organophosphates are used.

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±4 µg) and then transferred to good water quality, control water (CW; pH 6.8±0.1; <14±2 µg Ali). Al accumulation on gills after 2 and 7 days of acid/Al exposure was 35.3±14.1 and 26.6±1.8 µg g(-1) dry weight, respectively. These elevated levels decreased 2 days post transfer to CW and remained higher than in control (CON; 5-10 µg 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±4.8 mM) and ACID2 (127.6±3.7 mM) 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.

Impact of parasites on salmon recruitment in the Northeast Atlantic Ocean.

Parasites may have large effects on host population dynamics, marine fisheries and conservation, but a clear elucidation of their impact is limited by a lack of ecosystem-scale experimental data. We conducted a meta-analysis of replicated manipulative field experiments concerning the influence of parasitism by crustaceans on the marine survival of Atlantic salmon (Salmo salar L.). The data include 24 trials in which tagged smolts (totalling 283 347 fish; 1996-2008) were released as paired control and parasiticide-treated groups into 10 areas of Ireland and Norway. All experimental fish were infection-free when released into freshwater, and a proportion of each group was recovered as adult recruits returning to coastal waters 1 or more years later. Treatment had a significant positive effect on survival to recruitment, with an overall effect size (odds ratio) of 1.29 that corresponds to an estimated loss of 39 per cent (95% CI: 18-55%) of adult salmon recruitment. The parasitic crustaceans were probably acquired during early marine migration in areas that host large aquaculture populations of domesticated salmon, which elevate local abundances of ectoparasitic copepods-particularly Lepeophtheirus salmonis. These results provide experimental evidence from a large marine ecosystem that parasites can have large impacts on fish recruitment, fisheries and conservation.

Effects of the fungicide azoxystrobin on Atlantic salmon (Salmo salar L.) smolt.

Atlantic salmon smolts were exposed to three doses of the fungicide azoxystrobin for 4 days, and physiological blood parameters and transcriptional effects in liver and muscle were evaluated in search for potential negative effects. Azoxystrobin exposure mediated up-regulation of catalase, MAPK1 and IGFBP1 in liver tissue. Catalase, transferrin, IGFBP1 and TNFR were up-regulated and CYP1A down-regulated in muscle tissue. Blood parameters glucose, hematocrit, pCO(2), HCO(3) and pH grouped together with transcripts levels of MnSOD, MAPK1, IGFBP1, MAP3K7 and GPx4 in liver of fish exposed to the highest azoxystrobin concentration (352 µgL(-1)) using principal component analysis (PCA). In muscle, the blood parameters glucose, hematocrit, pCO(2), HCO(3) and pH grouped together with transcript levels of heme oxygenase, thioredoxin, MnSOD, TNFR and MMP9. These results suggest that the fungicide azoxystrobin affects mitochondrial respiration and mechanisms controlling cell growth and proliferation in fish and may have negative effects on juvenile Atlantic salmon.

Effects of acidic water and aluminum exposure on gill Na(+), K(+)-ATPase alpha-subunit isoforms, enzyme activity, physiology and return rates in Atlantic salmon (Salmo salar L.).

Na(+), K(+)-ATPase (NKA) is involved, through its role as a major driving force for electrochemical gradients, in a range of transmembrane transport processes. Maintenance of homeostasis in anadromous salmonids requires modulation of several gill ion secretory proteins as part of the preparatory adaptation and acclimation to marine life. Atlantic salmon smolts were exposed to combinations of low pH and inorganic aluminum (acid/Al(i)) in freshwater (FW) and were then transferred to seawater (SW) for studies of post-smolt performance. Gill mRNA levels of four NKA-alpha isoforms (alpha1a, alpha1b, alpha1c and alpha3) of the catalytic NKA subunit and NKA enzyme activity were measured. Moderate acid/Al treatment (MOD, pH 5.9+/-0.3, 15+/-9microgl(-1)Al(i)) prevented the FW preparatory increase in NKA activity observed in control (CON, pH 6.9+/-0.1, 8+/-3microgl(-1)Al(i)) smolts, while high acid/Al treatment (SEV, pH 5.6+/-0.2, 30+/-7microgl(-1)Al(i)) caused a rapid and persistent reduction in NKA activity. Correspondingly, a 3.3-fold increase in plasma glucose levels in the SEV groups concurrent with a decrease in plasma chloride levels suggest that acid/Al exposed fish were stressed and experienced problems maintaining ion homeostasis. Gill NKA activities in acid/Al exposed groups were re-established after 28 days in SW. Both long (9 days) and short-term (2.5 days) treatments had significant impact on isoform-specific Na(+), K(+)-ATPase alpha-subunit mRNA abundance in the FW period. Acid/Al exposed groups lacked the preparatory increases in all NKA-alpha isoform mRNA levels seen in the CON group, except for alpha1a. In contrast to the other isoforms measured, alpha1a mRNA abundance decreased sharply upon SW transfer, supporting the hypothesis of isozyme shifting as a mechanism of altering the gill from an ion absorbing to an ion excreting tissue during smoltification and SW exposure. Adult return rates to the Imsa river were significantly reduced both in short-term (78% of controls) and long-term (55% of controls) acid/Al exposures, emphasising the physiological and ecological consequences of acid/Al exposure during smoltification.