Effects of regional coordination of salmon louse control in reducing negative impacts of salmonid aquaculture on wild salmonids.

Parasitic salmon lice (Lepeophtheirus salmonis) are a constraint to the sustainable growth of salmonids in open net pens, and this issue has caused production to level off in recent years in the most aquaculture-intensive areas of Norway. The maximum allowed biomass at a regional level is regulated by using the so-called "traffic light" system, where salmon louse-induced mortality of migrating wild salmon post-smolts is evaluated against set targets. As a case study, we have investigated how a specific aquaculture-intensive area can reduce its louse levels sufficiently to achieve a low impact on wild salmon. Analyses of the output from a virtual post-smolt model that uses data on the reported number of salmon lice in fish farms as key input data and estimates the salmon louse-induced mortality of wild out-migrating Atlantic salmon post-smolts, suggested that female louse abundance on the local farms must be halved in spring to reach the goal implied by the traffic light system. The outcome of a modelling scenario simulating a proposed new plan for coordinated production and fallowing proved beneficial, with an overall reduction in louse infestations and treatment efforts. The interannual variability in louse abundance in spring, however, increased for this scenario, implying unacceptably high louse abundance when many farms were in their second production year. We then combined the scenario with coordinated production with other louse control measures. Only measures that reduced the density of farmed salmonids in open cages in the study area resulted in reductions in salmon louse infestations to acceptable levels. This could be achieved either by stocking with larger fish to reduce exposure time or by reducing fish numbers, e.g. by producing in closed units.

Lufenuron treatment temporarily represses gene expression and affects the SUMO pathway in liver of Atlantic salmon.

Lufenuron is a benzoylurea insecticide currently in use to combat sea lice infestation in salmon aquaculture in Chile. With pending approval in Norway, the aim of this work was to study the uptake and toxicity of lufenuron in liver tissue of Atlantic salmon. Juvenile salmon weighing 40 g were given a standard 7-day oral dose, and bioaccumulation and transcriptional responses in the liver were examined 1 day after the end-of-treatment (day 8) and after 1 week of elimination (day 14). Bioaccumulation levels of lufenuron were 29 ± 3 mg/kg at day 8 and 14 ± 1 mg/kg at day 14, indicating relatively rapid clearance. However, residues of lufenuron were still present in the liver after 513 days of depuration. The exposure gave a transient inhibition of transcription in the liver at day 8 (2437 significant DEGs, p-adj < .05), followed by a weaker compensatory response at day 14 (169 significant DEGs). Pathways associated with RNA metabolism such as the sumoylation pathway were most strongly affected at day 8, while the apelin pathway was most profoundly affected at day 14. In conclusion, this study shows that lufenuron easily bioaccumulates and that a standard 7-day oral dose induces a transient inhibition of transcription in liver of salmon.

The economic impact of decreased mortality and increased growth associated with preventing, replacing or improving current methods for delousing farmed Atlantic salmon in Norway.

Impacts of salmon lice is a major concern for a sustainable production of farmed Atlantic salmon in Norway. Most treatment methods for removal of salmon lice have associated increased mortality and decreased growth in a period after delousing, which affects the profitability of the farmer, and causes poor welfare and sustainability. In addition, the variance in mortality and growth, especially after non-medicinal treatment methods, is high, which makes it hard for a farmer to decide which control measure to apply to keep lice levels below the legal limit. In this study, we have applied a stochastic partial budget approach to assess the economic impact of reducing mortality and increasing growth of farmed Atlantic salmon by preventing, replacing and improving current delousing methods in Norway. We have simulated a production cycle of two different smolt-groups to find the outcome (harvested biomass, average end weight of the salmon, number of dead fish and feed consumption) of production cycles without or with two, three or four delousing treatments in the on-growing phase at sea. The results suggest that accounting for the biological losses associated with lice treatments is important when making choices of delousing strategies. The biological costs of increased mortality and decreased growth associated with especially non-medicinal treatments are expected to be high, but varies substantially. Therefore, the economic benefit of preventing or improving can also be high. The calculations imply that salmon producers could invest a considerable amount in measures for prevention or improvement of thermal treatments before break-even. For example could a farmer use on average 535,313 €/cage/ 1-yearling production in measure to prevent four thermal treatments before it is no longer economical beneficial. Depending on the performance of the four thermal treatments a farmer could use from 319,196-737,934 €/cage/ 1-yearling production on measures of improvement. Replacing one thermal treatment with another immediate treatment method has a minor economic benefit. The results further shows that sales value and feed consumption constitutes the largest share of the change in profit between different treatment regimes. The results from this study also show that not taking into account the risk of mortality and reduced growth associated with the different treatment methods of delousing, could lead to underestimating the benefit of improving, preventing and replacing treatments.

Policies to promote breeding for lice-resistant salmon: Incentives designed for resilient and sustainable growth in aquaculture.

Sea lice represent a persistent and growing problem, challenging the resilience and growth of the salmon aquaculture industry. In this Norwegian case study, we studied and discuss how the absence of policy instruments directed at stimulating breeding for lice resistance (LR) might be explained. We found well-documented opportunities for selection progress for LR. Hence, breeding on LR appears with an untapped potential. We discuss how market-based, legal, institutional and interest-based factors can explain the absence of policy instruments stimulating LR breeding. Methodologically, we obtained data from document and literature studies and interviews with key players (salmon breeders, farmers, nongovernmental organizations (NGOs) and governmental bodies in Norway). First, LR is a polygenic trait, which makes it poorly suited for patenting. Furthermore, if only a small proportion of fish farmers choose seeds with higher LR, other operators can easily take on the free-rider role because they will not suffer from reduced gain in growth performance as a result of a much stronger emphasis on LR in the breeding goal. The market is thus not expected to stimulate stronger selection for LR in Norwegian salmon breeding. Second, neither genetic engineering (e.g., gene editing), still struggling with consumer acceptance, nor the uncertainty associated with possible changes in the Norwegian Gene Technology Act stimulate investment in LR via, for example, CRISPR technology. Thirdly, public policy instruments in their entirety have targeted other types of innovations against salmon lice, and none have so far been used to stimulate breeding companies to emphasize LR more strongly in their breeding programmes. From a political point of view, it seems that breeding has been left to the market and the private sector. However, neither the NGOs nor the public seem to be aware of, or pay significant attention to, the breeding potential to improve LR and fish welfare. Fragmented management of the aquaculture sector can camouflage the close ties between political and business interests. The industry is hesitant to invest significantly in long-term breeding targets such as significantly higher genetic LR. This may strengthen the assumption that strong economic interests will reduce the role of science in knowledge-based management. As farmed salmon are increasingly being exposed to stressful delousing treatments, mortality and associated welfare problems have increased significantly. For instance, large fish die more often from cardiomyopathy syndrome (CMS), resulting in growing demand for CMS-resistant salmon. This gives rise to a paradoxical situation: increasing treatments with high mortality and fish welfare issues in farmed salmon, while the lice threat to wild salmon persists.

The skin mucosal barrier of lumpfish (Cyclopterus lumpus L.) is weakened by exposure to potential aquaculture production-related stressors.

Various cleaner fish species, such as the lumpfish (Cyclopterus lumpus L.), are used in the sea cage production of Atlantic salmon (Salmo salar L.) as a control measure against the ectoparasitic salmon louse (Lepeophtheirus salmonis). Nonetheless, during severe lice infestation, alternative treatments are required to control parasitic burden. The aim of this study was to gain insight into how lumpfish skin responds to different chemicals used to treat parasites. The authors collected skin from lumpfish from both research facilities (tank-reared fish) and commercial production (cage-reared fish) and used operational welfare indicators, in vitro models, histology and transcriptomics to study how the skin responded to two anti-parasitic oxidative chemicals, hydrogen peroxide (H2 O2 ) and peracetic acid. Lumpfish sampled from the farm were classified as clinically healthy or weak based on their morbidity status, and fish from each category were used to gain insight into how the therapeutics affect the skin barrier. Differences between healthy and weakened (moribund) fish, and between treated fish from each of the two groups, were observed. Histological examination showed an overall reduced skin quality in fish characterized as moribund, including different grades of exposed bony plates. In vitro oxidant-treated lumpfish skin had reduced the migration capacity of keratocytes, a weakened epidermal barrier, and altered gene transcription, changes that are known predisposing factors to secondary infections. Skin from non-treated, healthy fish sampled from commercial farms exhibited similar features and attributes to oxidant-exposed tank-reared fish from a research facility, suggesting that apparently healthy cage-held lumpfish exhibited stress responses in the epidermal barrier. The results of the study outline the risks and consequences lumpfish can face if accidentally subjected to potential anti-parasitic oxidant treatments aimed at Atlantic salmon. It also strengthens the evidence behind the requirement that lumpfish should be removed from the cages before being potentially exposed to this type of treatment and outlines the potential risks of differing husbandry practices upon lumpfish health, welfare and resilience.

The effect of different intensities of the ectoparasitic salmon lice (Lepeophtheirus salmonis) on Atlantic salmon (Salmo salar).

The effect of different intensities of the ectoparasitic salmon lice (Lepeophtheirus salmonis) on stress, growth and the expression of immune and wound healing transcripts in the skin of Atlantic salmon (Salmo salar) was investigated. Lice infection success and survival were similar at the chalimus and preadult stage in the low and high dose group, but infection success and survival were significantly lower in the high than in the low dose group at the adult stage. The expression of investigated transcripts was not correlated to lice intensities, but several of them were significantly differently expressed locally in the skin at the site of lice attachment in infected fish compared to controls. This included an up-regulation of pro-inflammatory markers at the site of lice attachment (e.g., interleukin 1-beta, interleukin 8 and the acute phase protein serum amyloid A), a reduction of markers of adaptive immunity (cluster of differentiation 8-alpha and immunoglobulin M) and decreased expression of the anti-inflammatory cytokine interleukin 10.

Time series covering up to four decades reveals major changes and drivers of marine growth and proportion of repeat spawners in an Atlantic salmon population.

Wild Atlantic salmon populations have declined in many regions and are affected by diverse natural and anthropogenic factors. To facilitate management guidelines, precise knowledge of mechanisms driving population changes in demographics and life history traits is needed.Our analyses were conducted on (a) age and growth data from scales of salmon caught by angling in the river Etneelva, Norway, covering smolt year classes from 1980 to 2018, (b) extensive sampling of the whole spawning run in the fish trap from 2013 onwards, and (c) time series of sea surface temperature, zooplankton biomass, and salmon lice infestation intensity.Marine growth during the first year at sea displayed a distinct stepwise decline across the four decades. Simultaneously, the population shifted from predominantly 1SW to 2SW salmon, and the proportion of repeat spawners increased from 3 to 7%. The latter observation is most evident in females and likely due to decreased marine exploitation. Female repeat spawners tended to be less catchable than males by anglers.Depending on the time period analyzed, marine growth rate during the first year at sea was both positively and negatively associated with sea surface temperature. Zooplankton biomass was positively associated with growth, while salmon lice infestation intensity was negatively associated with growth.Collectively, these results are likely to be linked with both changes in oceanic conditions and harvest regimes. Our conflicting results regarding the influence of sea surface temperature on marine growth are likely to be caused by long-term increases in temperature, which may have triggered (or coincided with) ecosystem shifts creating generally poorer growth conditions over time, but within shorter datasets warmer years gave generally higher growth. We encourage management authorities to expand the use of permanently monitored reference rivers with complete trapping facilities, like the river Etneelva, generating valuable long-term data for future analyses.

IIb-RAD-sequencing coupled with random forest classification indicates regional population structuring and sex-specific differentiation in salmon lice (Lepeophtheirus salmonis).

The aquaculture industry has been dealing with salmon lice problems forming serious threats to salmonid farming. Several treatment approaches have been used to control the parasite. Treatment effectiveness must be optimized, and the systematic genetic differences between subpopulations must be studied to monitor louse species and enhance targeted control measures. We have used IIb-RAD sequencing in tandem with a random forest classification algorithm to detect the regional genetic structure of the Norwegian salmon lice and identify important markers for sex differentiation of this species. We identified 19,428 single nucleotide polymorphisms (SNPs) from 95 individuals of salmon lice. These SNPs, however, were not able to distinguish the differential structure of lice populations. Using the random forest algorithm, we selected 91 SNPs important for geographical classification and 14 SNPs important for sex classification. The geographically important SNP data substantially improved the genetic understanding of the population structure and classified regional demographic clusters along the Norwegian coast. We also uncovered SNP markers that could help determine the sex of the salmon louse. A large portion of the SNPs identified to be under directional selection was also ranked highly important by random forest. According to our findings, there is a regional population structure of salmon lice associated with the geographical location along the Norwegian coastline.

Temperature affects settlement success of ectoparasitic salmon lice (Lepeophtheirus salmonis) and impacts the immune and stress response of Atlantic salmon (Salmo salar).

In this study, the effect of temperature on Atlantic salmon (Salmo salar) stress and immune response to the ectoparasitic salmon lice (Lepeophtheirus salmonis) was investigated. We found that infestation affected the expression of several immune and wound healing transcripts in the skin especially at the site of lice attachment compared to un-infested control fish. Moreover, expression patterns in the skin of infested fish suggest that host immune responses towards salmon lice are impaired at low temperatures. However, reduced lice infestation success and survival at the lowest investigated temperatures suggest that cold water temperatures are more detrimental to the lice than their fish hosts. Finally, temperature affected the stress response of the fish and infected fish had a higher increase in cortisol levels in response to handling (a stressor) than un-infested controls.

Energetic costs of ectoparasite infection in Atlantic salmon.

Parasites are widespread in nature, where they affect the energy budget of hosts, and depending on the imposed pathogenic severity, this may reduce host fitness. However, the energetic costs of parasite infections are rarely quantified. In this study, we measured metabolic rates in recently seawater adapted Atlantic salmon (Salmo salar) infected with the ectoparasitic copepod Lepeophtheirus salmonis and used an aerobic scope framework to assess the potential ecological impact of this parasite-host interaction. The early chalimus stages of L. salmonis did not affect either standard or maximum metabolic rates. However, the later mobile pre-adult stages caused an increase in both standard and maximum metabolic rate yielding a preserved aerobic scope. Notably, standard metabolic rates were elevated by 26%, presumably caused by increased osmoregulatory burdens and costs of mobilizing immune responses. The positive impact on maximum metabolic rates was unexpected and suggests that fish are able to transiently overcompensate energy production to endure the burden of parasites and thus allow for continuation of normal activities. However, infected fish are known to suffer reduced growth, and this suggests that a trade-off exists in acquisition and assimilation of resources despite an uncompromised aerobic scope. As such, when assessing impacts of environmental or biotic factors, we suggest that elevated routine costs may be a stronger predictor of reduced fitness than the available aerobic scope. Furthermore, studying the effects on parasitized fish in an ecophysiological context deserves more attention, especially considering interacting effects of other stressors in the Anthropocene.

Genomic analysis of the carboxylesterase family in the salmon louse (Lepeophtheirus salmonis).

The pyrethroid deltamethrin and the macrocyclic lactone emamectin benzoate (EMB) are used to treat infestations of farmed salmon by parasitic salmon lice, Lepeophtheirus salmonis. While the efficacy of both compounds against Atlantic populations of the parasite has decreased as a result of the evolution of resistance, the molecular mechanisms of drug resistance in L. salmonis are currently not fully understood. The functionally diverse carboxylesterases (CaE) family includes members involved in pesticide resistance phenotypes of terrestrial arthropods. The present study had the objective to characterize the CaE family in L. salmonis and assess its role in drug resistance. L. salmonis CaE homologues were identified by homology searches in the parasite's transcriptome and genome. The transcript expression of CaEs predicted to be catalytically competent was studied using quantitative reverse-transcription PCR in drug susceptible and multi-resistant L. salmonis. The above strategy led to the identification of 21 CaEs genes/pseudogenes. Phylogenetic analyses assigned 13 CaEs to clades involved in neurodevelopmental signaling and cell adhesion, while three sequences were predicted to encode secreted enzymes. Ten CaEs were identified as being potentially catalytically competent. Transcript expression of acetylcholinesterase (ace1b) was significantly increased in multi-resistant lice compared to drug-susceptible L. salmonis, with transcript abundance further increased in preadult-II females following EMB exposure. In summary, results from the present study demonstrate that L. salmonis possesses fewer CaE gene family members than most arthropods characterized so far. Drug resistance in L. salmonis was associated with overexpression of ace1b.

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.

Responses to Mineral Supplementation and Salmon Lice (Lepeophtheirus salmonis) Infestation in Skin Layers of Atlantic Salmon (Salmo salar L.).

The crustacean ectoparasite salmon louse (Lepeophtheirus salmonis), which severely affects Atlantic salmon health and welfare is one of the main problems of commercial aquaculture. In the present study, fish were fed a diet supplemented with extra minerals through the inclusion of a commercial additive (Biofeed Forte Salmon), substituting wheat in the control diet, before experimental infestation with salmon lice. Lice counts reduced with time but with no apparent effect of the diets. Further, fish fed the mineral diet had an overall higher number of blue (acidic) mucous cells, while the ratio of purple mucous cells was higher in the mineral diet. The transcriptional response in skin was enhanced at 7 dpc (copepodite life stage) in fish fed the mineral diet including immune and stress responses, while at 21 dpc (pre-adult life stage), the difference disappeared, or reversed with stronger induction in the control diet. Overall, 9.3% of the genes affected with lice also responded to the feed, with marked differences in outer (scale + epidermis) and inner (dermis) skin layers. A comparison of transcriptome data with five datasets from previous trials revealed common features and gene markers of responses to lice, stress, and mechanically induced wounds. Results suggested a prevalence of generic responses in wounded skin and lice-infected salmon.

Estimating cage-level mortality distributions following different delousing treatments of Atlantic salmon (salmo salar) in Norway.

This retrospective descriptive study estimates cage-level mortality distributions after six immediate delousing methods: thermal, mechanical, hydrogen peroxide, medicinal, freshwater and combination of medicinal treatments. We investigated mortality patterns associated with 4 644 delousing treatment of 1 837 cohorts of farmed Atlantic salmon (Salmo salar) stocked in sea along the Norwegian coast between 2014 and 2017. The mortality is expressed as mortality rates. We found distributions of delta mortality rate within 1, 7 and 14 days after all six delousing treatments, using mortality rate within 7 days before treatments as baseline. The results show that we can expect increased mortality rates after all six delousing methods. The median delta mortality rates after thermal and mechanical delousing are 5.4 and 6.3 times higher than medicinal treatment, respectively, for the 2017 year-class. There is a reduction in the delta median mortality for thermal and freshwater delousing from 2015 to 2019. There is a wide variability in the mortality rates, in particular for thermal delousing. Our results suggest that the variability in delta mortality for thermal delousing has been reduced from the 2014 to 2017 year-class, indicating an improvement of the technique. However, a significant increase in the number of thermal treatments from 14 in 2015 to 738 in 2018 probably contributes to the overall increased mortality in Norwegian salmon farming.

Bias in self-reported parasite data from the salmon farming industry.

Many industries are required to monitor themselves in meeting regulatory policies intended to protect the environment. Self-reporting of environmental performance can place the cost of monitoring on companies rather than taxpayers, but there are obvious risks of bias, often addressed through external audits or inspections. Surprisingly, there have been relatively few empirical analyses of bias in industry self-reported data. Here, we test for bias in reporting of environmental compliance data using a unique data set from Canadian salmon farms, where companies monitor the number of parasitic sea lice on fish in open sea pens, in order to minimize impacts on wild fish in surrounding waters. We fit a hierarchical population-dynamics model to these sea-louse count data using a Bayesian approach. We found that the industry's monthly counts of two sea-louse species, Caligus clemensi and Lepeophtheirus salmonis, increased by a factor of 1.95 (95% credible interval: 1.57, 2.42) and 1.18 (1.06, 1.31), respectively, in months when counts were audited by the federal fisheries department. Consequently, industry sea-louse counts are less likely to trigger costly but mandated delousing treatments intended to avoid sea-louse epidemics in wild juvenile salmon. These results highlight the potential for combining external audits of industry self-reported data with analyses of their reporting to maintain compliance with regulations, achieve intended conservation goals, and build public confidence in the process.

Protective Immunization of Atlantic Salmon (Salmo salar L.) against Salmon Lice (Lepeophtheirus salmonis) Infestation.

Vaccination against salmon lice (Lepeophtheirus salmonis) is a means of control that averts the negative effects of chemical approaches. Here, we studied the immunogenicity and protective effect of a vaccine formulation (based on a salmon lice-gut recombinant protein [P33]) against Lepeophtheirus salmonis infestation in Atlantic salmon in a laboratory-based trial. Our findings revealed that P33 vaccine can provide a measure of protection against immature and adult salmon lice infestation. This protection seemed to be vaccine dose-dependent, where higher doses resulted in lower parasitic infestation rates. We also provide immunological evidence confirming that P33-specific immune response can be triggered in Atlantic salmon after P33 vaccination, and that production of P33-specific antibodies in blood can be detected in vaccinated fish. The negative correlation between P33-specific IgM in salmon plasma and salmon lice numbers on vaccinated fish suggests that protection against lice can be mediated by the specific antibody in salmon plasma. The success of P33 vaccination in protecting salmon against lice confirms the possibility of employing the hematophagous nature of the parasite to deliver salmon-specific antibodies against lice-gut proteins.

Investigation of deltamethrin resistance in salmon lice (Lepeophtheirus salmonis) provides no evidence for roles of mutations in voltage-gated sodium channels.

BACKGROUND: The pyrethroid deltamethrin is used to treat infestations of farmed salmon by parasitic salmon lice, Lepeophtheirus salmonis (Krøyer). However, the efficacy of deltamethrin for salmon delousing is threatened by resistance development. In terrestrial arthropods, knockdown resistance (kdr) mutations of the voltage-gated sodium channel (Nav ), the molecular target for pyrethroids, can cause deltamethrin resistance. A putative kdr mutation of an L. salmonis sodium channel homologue (LsNav 1.3 I936V) has been identified previously. At the same time, deltamethrin resistance of L. salmonis has been shown to be inherited maternally and to be associated with mitochondrial DNA (mtDNA) mutations. This study assessed potential roles of the above putative kdr mutation as a determinant of deltamethrin resistance in laboratory strains and field populations of L. salmonis. RESULTS: The deltamethrin-resistant L. salmonis strain IoA-02 expresses the LsNav 1.3 I936V mutation but was susceptible to the non-ester pyrethroid etofenprox, a compound against which pyrethroid-resistant arthropods are usually cross-resistant if resistance is caused by Nav mutations. In a family derived from a cross between an IoA-02 male and a drug-susceptible female lacking the kdr mutation, deltamethrin resistance was not associated with the genotype at the LsNav 1.3 locus (P > 0.05). Similarly, in Scottish field populations of L. salmonis, LsNav 1.3 I936V showed no association with deltamethrin resistance. By contrast, genotypes at the mtDNA loci A14013G and A9030G were significantly associated with deltamethrin resistance (P < 0.001). CONCLUSION: In the studied L. salmonis isolates, deltamethrin resistance was unrelated to the LsNav 1.3 I936V mutation, but showed close association with mtDNA mutations.

The effects of teflubenzuron on mortality, physiology and accumulation in Capitella sp.

The chitin synthesis inhibitor teflubenzuron (TFB) is a feed antiparasitic agents used to impede molting of the salmon lice, an ecto-parasite that severely affects the salmon industry. Low absorption of oral administered TFB may cause elevated concentrations in the feces discharged from the salmon into the benthic environment. The polychaete Capitella sp. are often dominant in such habitats and consume organic waste deposited on the sediment. In the present study, Capitella sp. were exposed to doses of TFB in salmon feed of 1, 2 and 4 g TFB kg-1 (0 g TFB kg-1 in control group) over an experimental period of 32 days. Cumulative mortality was 12%-15% in both treatment groups with 1 and 2 g TFB kg-1 and reached 27% in the group with 4 g TFB kg-1. Only the highest dose (4 g TFB kg-1) negatively affected feed intake, growth and respiration of the polychaetes while food conversion efficiency was not affected. At the end of the experiment, the concentrations of TFB in the Capitella sp. were high, in the range of 9.24-10.32 µg g-1 for the three treatment groups. It was suggested that a maximum level of absorption rate was reached, also for the lowest dose. High concentrations of TFB in the Capitella sp. might pose a risk to crustaceans that forage for polychaetes in the vicinity of fish farms. We conclude that the effects of TFB on Capitella sp. may therefore primarily be to the predators rather than the Capitella sp.

Identification of critical enzymes in the salmon louse chitin synthesis pathway as revealed by RNA interference-mediated abrogation of infectivity.

Treatment of infestation by the ectoparasite Lepeophtheirus salmonis relies on a small number of chemotherapeutant treatments that currently meet with limited success. Drugs targeting chitin synthesis have been largely successful against terrestrial parasites where the pathway is well characterised. However, a comparable approach against salmon lice has been, until recently, less successful, likely due to a poor understanding of the chitin synthesis pathway. Post-transcriptional silencing of genes by RNA interference (RNAi) is a powerful method for evaluation of protein function in non-model organisms and has been successfully applied to the salmon louse. In the present study, putative genes coding for enzymes involved in L. salmonis chitin synthesis were characterised after knockdown by RNAi. Nauplii I stage L. salmonis were exposed to double-stranded (ds) RNA specific for several putative non-redundant points in the pathway: glutamine: fructose-6-phosphate aminotransferase (LsGFAT), UDP-N-acetylglucosamine pyrophosphorylase (LsUAP), N-acetylglucosamine phosphate mutase (LsAGM), chitin synthase 1 (LsCHS1), and chitin synthase 2 (LsCHS2). Additionally, we targeted three putative chitin deacetylases (LsCDA4557, 5169 and 5956) by knockdown. Successful knockdown was determined after moulting to the copepodite stage by real-time quantitative PCR (RT-qPCR), while infectivity potential (the number of attached chalimus II compared with the initial number of larvae in the system) was measured after exposure to Atlantic salmon and subsequent development on their host. Compared with controls, infectivity potential was not compromised in dsAGM, dsCHS2, dsCDA4557, or dsCDA5169 groups. In contrast, there was a significant effect in the dsUAP-treated group. However, of most interest was the treatment with dsGFAT, dsCHS1, dsCHS1+2, and dsCDA5956, which resulted in complete abrogation of infectivity, despite apparent compensatory mechanisms in the chitin synthesis pathway as detected by qPCR. There appeared to be a common phenotypic effect in these groups, characterised by significant aberrations in appendage morphology and an inability to swim. Ultrastructurally, dsGFAT showed a significantly distorted procuticle without distinct exo/endocuticle and intermittent electron dense (i.e. chitin) inclusions, and together with dsUAP and dsCHS1, indicated delayed entry to the pre-moult phase.

Parasites under pressure: salmon lice have the capacity to adapt to depth-based preventions in aquaculture.

The evolution of pesticide resistance has driven renewed interest in non-chemical pest controls in agriculture. Spatial manipulations (physical barriers and fallowing, for example) can be an effective method of prevention, but these too might impose selection and cause rapid adaptation in pests. In salmon aquaculture, various non-chemical approaches have emerged to combat parasitic salmon lice (Lepeophtheirus salmonis) - a major pest with clear signs of evolved chemical resistance. 'Depth-based' preventions, now widely implemented, reduce infestation rates by physically segregating salmon from lice in their infective copepodid stage occurring in surface waters. Copepodids distributed deeper in the water column, however, can bypass these barriers and infest farms. If swimming depth is a heritable trait, we may see rapid evolutionary shifts in response to widespread depth-based prevention. We collected lice from Norwegian salmon farms and assayed more than 11,250 of their laboratory-reared offspring across 37 families. The vertical distributions of copepodids were measured using experimental water columns pressurised to simulate conditions at 0, 5 and 10 m depths. We demonstrated that lice respond strongly to hydrostatic pressure: an increase in pressure doubled the number of lice that migrated to the top of columns. There was also a large effect of family on this response, with the percentage of lice ascending to the top of pressurised columns ranging from 17 to 79% across families. Families with a weak swimming response to pressure are expected to occur deeper in the water column and so be more likely to infest farms employing depth-based preventions. If this between-family variation reflects genetic variation, then the parasite population may have the capacity to adapt to preventative measures. Such adaptation would have important commercial and ecological implications.