Welfare and performance of ballan wrasse (Labrus bergylta) reared at two different temperatures after a preparatory feeding trial with enhanced dietary eicosapentaenoic acid.

Concerns have long been raised about the welfare of ballan wrasse (Labrus bergylta) used for the biological control of sea lice in Atlantic salmon (Salmo salar) aquaculture. This study assessed the effect of increased dietary eicosapentaenoic acid (EPA) levels and initial condition factor (CF) on the subsequent performance and welfare of ballan wrasse farmed in high and low water temperatures. Fish were fed a diet with either commercial or high EPA levels for 3 months at 15°C. Subsequently, fish were tagged with a passive integrated transponder, measured for their CF and divided into two groups consisting of fish from both treatments and reared for 4.5 months at either 15 or 6°C fed a commercial diet. Each fish was categorized as high (≥2.7) or low CF (<2.7) fish based on the calculated average CF of the population. Dietary composition influenced the fatty acid (FA) profile of the stored lipids without affecting the growth or welfare of ballan wrasse. Fish reared at 15°C showed higher growth, more fat and energy reserves and less ash content. Fish reared at 6°C lost weight, using up their body lipids at the end of the temperature trial. Gene expression analyses showed upregulation of the positive growth marker (GHrα) and two genes involved in the synthesis and oxidation of FAs (elovl5, cpt1) and downregulation of the negative growth marker (mstn) in fish reared at 15°C compared to those reared at 6°C. Fish reared at 6°C showed upregulated levels of il-6 compared to those reared at 15°C, suggesting an enhanced immune reaction in response to low temperature. Fish with high CF showed better survival, growth and performance compared to those with low CF. External welfare scoring showed higher prevalence and severity in emaciation, scale loss and the sum index score (of all measured welfare parameters) in fish reared at 6°C compared to those reared at 15°C and better welfare in fish with high CF compared to those with low CF. Histological examination of the skin showed that fish reared at 6°C had decreased epidermal thickness, a lower overall number of mucous cells in the inner and outer epidermis and a different organization of mucous cells compared to fish reared at 15°C, indicating stress in fish reared at 6°C. Overall, low water temperatures had profound effects on the performance and external and internal welfare parameters of ballan wrasse and can be considered a stressor likely affecting the delousing efficacy. These findings support the seasonal use of different cleaner fish species. High CF, but not increased dietary EPA levels, appeared to help fish cope better with low water temperatures and should thus be assessed and considered before deploying them in salmon cages.

Applying genetic technologies to combat infectious diseases in aquaculture.

Disease and parasitism cause major welfare, environmental and economic concerns for global aquaculture. In this review, we examine the status and potential of technologies that exploit genetic variation in host resistance to tackle this problem. We argue that there is an urgent need to improve understanding of the genetic mechanisms involved, leading to the development of tools that can be applied to boost host resistance and reduce the disease burden. We draw on two pressing global disease problems as case studies-sea lice infestations in salmonids and white spot syndrome in shrimp. We review how the latest genetic technologies can be capitalised upon to determine the mechanisms underlying inter- and intra-species variation in pathogen/parasite resistance, and how the derived knowledge could be applied to boost disease resistance using selective breeding, gene editing and/or with targeted feed treatments and vaccines. Gene editing brings novel opportunities, but also implementation and dissemination challenges, and necessitates new protocols to integrate the technology into aquaculture breeding programmes. There is also an ongoing need to minimise risks of disease agents evolving to overcome genetic improvements to host resistance, and insights from epidemiological and evolutionary models of pathogen infestation in wild and cultured host populations are explored. Ethical issues around the different approaches for achieving genetic resistance are discussed. Application of genetic technologies and approaches has potential to improve fundamental knowledge of mechanisms affecting genetic resistance and provide effective pathways for implementation that could lead to more resistant aquaculture stocks, transforming global aquaculture.

Quantitative Genetics of Smoltification Status at the Time of Seawater Transfer in Atlantic Salmon (Salmo Salar).

High mortality during grow out in the sea is a challenge for farmed Atlantic salmon production in Norway and globally, which is partly attributed to suboptimal smolt quality. In this study, two groups of pre-smolts were put on a standard light smoltification regime with alternating 12L:12D per day for 6 weeks (Phase I), followed by 24L:0D per day for 6 weeks (Phase II); one group was 0 + smolt (EXP1) and the other 1 + smolt (EXP2). To monitor the smoltification status of the fish, 100 (EXP1) and 60 (EXP2) fish were randomly sampled per week during Phase II. The following phenotypes for smoltification status were studied: RT-qPCR relative mRNA expression of values of two alpha catalytic subunits of the variants of the Na+K+ATPase (NKA) expressed in the sampled gill tissues of each fish. The first variant, alpha1a with increased expression in freshwater (FW) and the second variant alpha1b with increased expression in seawater variant (SW), as well as their ratio SW/FW. At the optimal time for seawater transfer based on the SW/FW trait, 1,000 (at sixth sampling of EXP1) and 1,500 (at fifth sampling of EXP2) fish were sampled for genetic parameter estimation. The individual variation in FW, SW, and SW/FW was very large at each of the seven samplings indicating a large variation among individuals in the optimum time of transfer to seawater. SW/FW showed significant genetic variation in both 0+ and 1+ smolts, which indicates the possibility for selection for improved synchronization of smoltification status of Atlantic salmon at the time where the largest proportion of the fish is considered to be smolt. However, the genetic correlation between SW/FW of 0+ and 1+ was not significantly different from zero indicating very little shared genetic variation in SW/FW in 0+ and 1+ fish. Smoltification phenotypes showed temporal progression over the smoltification period, and this progression varied between 0+ and 1+ smolt highlighting the importance of correctly timing the major sampling point, and when cohorts are transferred to seawater. This also highlighted the need for further research into noninvasive methods of objectively measuring individual smoltification through time and subsequent smolt survival and growth rate at sea.

Quantitative Genetics of Growth Rate and Filet Quality Traits in Atlantic Salmon Inferred From a Longitudinal Bayesian Model for the Left-Censored Gaussian Trait Growth Rate.

In selective breeding programs for Atlantic salmon, test fish are slaughtered at an average body weight where growth rate and carcass traits as filet fat (F F), filet pigment (F P) and visceral fat index (F F) are recorded. The objective of this study was to obtain estimates of genetic correlations between growth rate (GR), and the three carcass quality traits when fish from the same 206 families (offspring of 120 sires and 206 dams from 2 year-classes) were recorded both at the same age (SA) and about the same body weight (SW). In the SW group, the largest fish were slaughtered at five different slaughter events and the remaining fish at the sixth slaughter event over 6 months. Estimates of genetic parameters for the traits were obtained from a Bayesian multivariate model for (potentially) truncated Gaussian traits through a Gibbs sampler procedure in which phantom GR values were obtained for the unslaughtered, and thus censored SW group fish at each slaughter event. The heritability estimates for the same trait in each group was similar; about 0.2 for FF, 0.15 for FP and 0.35 for VF and GR. The genetic correlation between the same traits in the two groups was high for growth rate (0.91 ± 0.05) visceral index (0.86 ± 0.05), medium for filet fat (0.45 ± 0.17) and low for filet pigment (0.13 ± 0.27). Within the two groups, the genetic correlation between growth rate and filet fat changed from positive (0.59 ± 0.14) for the SA group to negative (-0.45 ± 0.17) for the SW group, while the genetic correlation between growth rate and filet pigment changed from negative (-0.33 ± 0.22) for the SA group to positive (0.62 ± 0.16) for the SW group. The genetic correlation of growth rate with FF and FP is sensitive to whether the latter traits are measured at the same age or the same body weight. The results indicate that selection for increased growth rate is not expected to have a detrimental effect on the quality traits if increased growth potential is realized through a reduced production time.

Genome-wide association mapping and accuracy of predictions for amoebic gill disease in Atlantic salmon (Salmo salar).

Amoebic gill disease (AGD) is a parasitic disease caused by the amoeba Paramoeba perurans, which colonizes the gill tissues and causes distress for the host. AGD can cause high morbidity and mortalities in salmonid and non-salmonid fish species. To understand the genetic basis of AGD and improve health status of farmed A. salmon, a population of ~ 6,100 individuals belonging to 150 full-sib families was monitored for development of AGD in the sea of Ireland. The population was followed for two rounds of AGD infections, and fish were gill scored to identify severity of disease in first (N = 3,663) and the second (N = 3,511) infection with freshwater treatment after the first gill-scoring. A subset of this gill-scored population (N = 1,141) from 119 full-sib families were genotyped with 57,184 SNPs using custom-made Affymetrix SNP-chip. GWAS analyses were performed which resulted in five significantly associated SNP variants distributed over chromosome 1, 2 and 5. Three candidate genes; c4, tnxb and slc44a4 were found within QTL region of chromosome 2. The tnxb and c4 genes are known to be a part of innate immune system, and may play a role in resistance to AGD. The gain in prediction accuracy obtained by involving genomic information was 9-17% higher than using traditional pedigree information.

QTLs Associated with Resistance to Cardiomyopathy Syndrome in Atlantic Salmon.

Cardiomyopathy syndrome (CMS) caused by piscine myocarditis virus is a major disease affecting the Norwegian Atlantic salmon industry. Three different populations of Atlantic salmon from the Mowi breeding program were used in this study. The first 2 populations (population 1 and 2) were naturally infected in a field outbreak, while the third population (population 3) went through a controlled challenged test. The aim of the study was to estimate the heritability, the genetic correlation between populations and perform genome-wide association analysis for resistance to this disease. Survival data from population 1 and 2 and heart atrium histology score data from population 3 was analyzed. A total of 571, 4312, and 901 fish from population 1, 2, and 3, respectively were genotyped with a noncommercial 55,735 Affymetrix marker panel. Genomic heritability ranged from 0.12 to 0.46 and the highest estimate was obtained from the challenge test dataset. The genetic correlation between populations was moderate (0.51-0.61). Two chromosomal regions (SSA27 and SSA12) contained single nucleotide polymorphisms associated with resistance to CMS. The highest association signal (P = 6.9751 × 10-27) was found on chromosome 27. Four genes with functional roles affecting viral resistance (magi1, pi4kb, bnip2, and ha1f) were found to map closely to the identified quantitative trait loci (QTLs). In conclusion, genetic variation for resistance to CMS was observed in all 3 populations. Two important quantitative trait loci were detected which together explain half of the total genetic variance, suggesting strong potential application for marker-assisted selection and genomic predictions to improve CMS resistance.

Genomic and Transcriptomic Analysis of Amoebic Gill Disease Resistance in Atlantic Salmon (Salmo salar L.).

Amoebic gill disease (AGD) is one of the most important parasitic diseases of farmed Atlantic salmon. It is a source of major economic loss to the industry and poses significant threats to animal welfare. Previous studies have shown that resistance against this disease has a moderate, heritable genetic component, although the genes and the genetic pathways that contribute to this process have yet to be elucidated. In this study, to identify the genetic mechanisms of AGD resistance, we first investigated the molecular signatures of AGD infection in Atlantic salmon through a challenge model, where we compared the transcriptome profiles of the naïve and infected animals. We then conducted a genome-wide association analysis with 1,333 challenged tested fish to map the AGD resistance genomic regions, supported by the results from the transcriptomic data. Further, we investigated the potential of incorporating gene expression analysis results in genomic prediction to improve prediction accuracy. Our data suggest thousands of genes have modified their expression following infection, with a significant increase in the transcription of genes with functional properties in cell adhesion and a sharp decline in the abundance of various components of the immune system genes. From the genome-wide association analysis, QTL regions on chromosomes ssa04, ssa09, and ssa13 were detected to be linked with AGD resistance. In particular, we found that QTL region on ssa04 harbors members of the cadherin gene family. These genes play a critical role in target recognition and cell adhesion. The QTL region on ssa09 also is associated with another member of the cadherin gene family, protocadherin Fat 4. The associated genetic markers on ssa13 span a large genomic region that includes interleukin-18-binding protein, a gene with function essential in inhibiting the proinflammatory effect of cytokine IL18. Incorporating gene expression information through a weighted genomic relationship matrix approach decreased genomic prediction accuracy and increased bias of prediction. Together, these findings help to improve our breeding programs and animal welfare against AGD and advance our knowledge of the genetic basis of host-pathogen interactions.

Genetics of Growth Reaction Norms in Farmed Rainbow Trout.

Rainbow trout is farmed globally under diverse uncontrollable environments. Fish with low macroenvironmental sensitivity (ES) of growth is important to thrive and grow under these uncontrollable environments. The ES may evolve as a correlated response to selection for growth in one environment when the genetic correlation between ES and growth is nonzero. The aims of this study were to quantify additive genetic variance for ES of body weight (BW), defined as the slope of reaction norm across breeding environment (BE) and production environment (PE), and to estimate the genetic correlation (rg(int, sl)) between BW and ES. To estimate heritable variance of ES, the coheritability of ES was derived using selection index theory. The BW records from 43,040 rainbow trout performing either in freshwater or seawater were analysed using a reaction norm model. High additive genetic variance for ES (9584) was observed, inferring that genetic changes in ES can be expected. The coheritability for ES was either -0.06 (intercept at PE) or -0.08 (intercept at BE), suggesting that BW observation in either PE or BE results in low accuracy of selection for ES. Yet, the rg(int, sl) was negative (-0.41 to -0.33) indicating that selection for BW in one environment is expected to result in more sensitive fish. To avoid an increase of ES while selecting for BW, it is possible to have equal genetic gain in BW in both environments so that ES is maintained stable.

Genetic (co)variance of rainbow trout (Oncorhynchus mykiss) body weight and its uniformity across production environments.

BACKGROUND: When rainbow trout from a single breeding program are introduced into various production environments, genotype-by-environment (GxE) interaction may occur. Although growth and its uniformity are two of the most important traits for trout producers worldwide, GxE interaction on uniformity of growth has not been studied. Our objectives were to quantify the genetic variance in body weight (BW) and its uniformity and the genetic correlation (rg) between these traits, and to investigate the degree of GxE interaction on uniformity of BW in breeding (BE) and production (PE) environments using double hierarchical generalized linear models. Log-transformed data were also used to investigate whether the genetic variance in uniformity of BW, GxE interaction on uniformity of BW, and rg between BW and its uniformity were influenced by a scale effect. RESULTS: Although heritability estimates for uniformity of BW were low and of similar magnitude in BE (0.014) and PE (0.012), the corresponding coefficients of genetic variation reached 19 and 21%, which indicated a high potential for response to selection. The genetic re-ranking for uniformity of BW (rg = 0.56) between BE and PE was moderate but greater after log-transformation, as expressed by the low rg (-0.08) between uniformity in BE and PE, which indicated independent genetic rankings for uniformity in the two environments when the scale effect was accounted for. The rg between BW and its uniformity were 0.30 for BE and 0.79 for PE but with log-transformed BW, these values switched to -0.83 and -0.62, respectively. CONCLUSIONS: Genetic variance exists for uniformity of BW in both environments but its low heritability implies that a large number of relatives are needed to reach even moderate accuracy of selection. GxE interaction on uniformity is present for both environments and sib-testing in PE is recommended when the aim is to improve uniformity across environments. Positive and negative rg between BW and its uniformity estimated with original and log-transformed BW data, respectively, indicate that increased BW is genetically associated with increased variance in BW but with a decrease in the coefficient of variation. Thus, the scale effect substantially influences the genetic parameters of uniformity, especially the sign and magnitude of its rg.

Survival, growth and sexual maturation in Atlantic salmon exposed to infectious pancreatic necrosis: a multi-variate mixture model approach.

BACKGROUND: Outbreaks of infectious pancreatic necrosis (IPN) in Atlantic salmon can result in reduced growth rates in a fraction of the surviving fish (runts). Genetic and environmental variation also affects growth rates within different categories of healthy animals and runts, which complicates identification of runts. Mixture models are commonly used to identify the underlying structures in such data, and the aim of this study was to develop Bayesian mixture models for the genetic analysis of health status (runt/healthy) of surviving fish from an IPN outbreak. METHODS: Five statistical models were tested on data consisting of 10 972 fish that died and 3959 survivors with recorded growth data. The most complex models (4 and 5) were multivariate normal-binary mixture models including growth, sexual maturity and field survival traits. Growth rate and liability of sexual maturation were treated as two-component normal mixtures, assuming phenotypes originated from two potentially overlapping distributions, (runt/normal). Runt status was an unobserved binary trait. These models were compared to mixture models with fewer traits (Models 2 and 3) and a classical linear animal model for growth (Model 1). RESULTS: Assuming growth as a mixture trait improved the predictive ability of the statistical model considerably (Model 2 vs. 1). The final models (4 and 5) yielded the following results: estimated (underlying) heritabilities were moderate for growth in healthy fish (0.32 ± 0.04 and 0.35 ± 0.05), runt status (0.39 ± 0.07 and 0.36 ± 0.08) and sexual maturation (0.33 ± 0.05), and high for field survival (0.47 ± 0.03 and 0.48 ± 0.03). Growth in healthy animals, runt status and survival showed consistent favourable genetic associations. Sexual maturation showed an unfavourable non-significant genetic correlation with runt status, but favourable genetic correlations with other traits. The estimated fraction of healthy fish was 81-85%. The estimated breeding values for runt status and (normal) growth were consistent for the most complex models (4 and 5), but showed imperfect correlations with estimated breeding values from the simpler models. CONCLUSIONS: Modelling growth in IPN survivors as a mixture trait improved the predictive ability of the model compared with a classical linear model. The results indicated considerable genetic variation in health status among survivors. Mixture modelling may be useful for the genetic analysis of diseases detected mainly through indicator traits.

Quantitative genetics of Taura syndrome resistance in Pacific white shrimp (Penaeus vannamei): a cure model approach.

BACKGROUND: In aquaculture breeding, resistance against infectious diseases is commonly assessed as time until death under exposure to a pathogen. For some diseases, a fraction of the individuals may appear as "cured" (non-susceptible), and the resulting survival time may thus be a result of two confounded underlying traits, i.e., endurance (individual hazard) and susceptibility (whether at risk or not), which may be accounted for by fitting a cure survival model. We applied a cure model to survival data of Pacific white shrimp (Penaeus vannamei) challenged with the Taura syndrome virus, which is one of the major pathogens of Panaeid shrimp species. METHODS: In total, 15,261 individuals of 513 full-sib families from three generations were challenge-tested in 21 separate tests (tanks). All challenge-tests were run until mortality naturally ceased. Time-until-event data were analyzed with a mixed cure survival model using Gibbs sampling, treating susceptibility and endurance as separate genetic traits. RESULTS: Overall mortality at the end of test was 28%, while 38% of the population was considered susceptible to the disease. The estimated underlying heritability was high for susceptibility (0.41 ± 0.07), but low for endurance (0.07 ± 0.03). Furthermore, endurance and susceptibility were distinct genetic traits (rg = 0.22 ± 0.25). Estimated breeding values for endurance and susceptibility were only moderately correlated (0.50), while estimated breeding values from classical models for analysis of challenge-test survival (ignoring the cured fraction) were closely correlated with estimated breeding values for susceptibility, but less correlated with estimated breeding values for endurance. CONCLUSIONS: For Taura syndrome resistance, endurance and susceptibility are apparently distinct genetic traits. However, genetic evaluation of susceptibility based on the cure model showed clear associations with standard genetic evaluations that ignore the cure fraction for these data. Using the current testing design, genetic variation in observed survival time and absolute survival at the end of test were most likely dominated by genetic variation in susceptibility. If the aim is to reduce susceptibility, earlier termination of the challenge-test or back-truncation of the follow-up period should be avoided, as this may shift focus of selection towards endurance rather than susceptibility.

Hepatic gene expression profiling reveals protective responses in Atlantic salmon vaccinated against furunculosis.

BACKGROUND: Furunculosis, a disease caused with gram negative bacteria Aeromonas salmonicida produces heavy losses in aquaculture. Vaccination against furunculosis reduces mortality of Atlantic salmon but fails to eradicate infection. Factors that determine high individual variation of vaccination efficiency remain unknown. We used gene expression analyses to search for the correlates of vaccine protection against furunculosis in Atlantic salmon. RESULTS: Naïve and vaccinated fish were challenged by co-habitance. Fish with symptoms of furunculosis at the onset of mass mortality (LR - low resistance) and survivors (HR - high resistance) were sampled. Hepatic gene expression was analyzed with microarray (SFA2.0 - immunochip) and real-time qPCR. Comparison of LR and HR indicated changes associated with the protection and results obtained with naïve fish were used to find and filter the vaccine-independent responses. Genes involved in recruitment and migration of immune cells changed expression in both directions with greater magnitude in LR. Induction of the regulators of immune responses was either equal (NFkB) or greater (Jun) in LR. Expression levels of proteasome components and extracellular proteases were higher in LR while protease inhibitors were up-regulated in HR. Differences in chaperones and protein adaptors, scavengers of reactive oxygen species and genes for proteins of iron metabolism suggested cellular and oxidative stress in LR. Reduced levels of free iron and heme can be predicted in LR by gene expression profiles with no protection against pathogen. The level of complement regulation was greater in HR, which showed up-regulation of the components of membrane attack complex and the complement proteins that protect the host against the auto-immune damages. HR fish was also characterized with up-regulation of genes for proteins involved in the protection of extracellular matrix, lipid metabolism and clearance of endogenous and exogenous toxic compounds. A number of genes with marked expression difference between HR and LR can be considered as positive and negative correlates of vaccine protection against furunculosis. CONCLUSION: Efficiency of vaccination against furunculosis depends largely on the ability of host to neutralize the negative impacts of immune responses combined with efficient clearance and prevention of tissue damages.