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.

Warm water treatment increased mortality risk in salmon.

Thermal treatment is a controversial method to control sea lice in the Atlantic salmon farming industry. This study aimed to complement the growing evidence base to document the impact of thermal treatments on salmon welfare, behaviour, physiology and health. Here, fish were treated two times (four weeks apart) for 30 s in either 27, 30, or 33 °C warm water, and parameters were compared to a procedural control (exposed to their holding temperature of 14 °C) or a negative control (where no treatments were applied). The fish had a clear behavioural response to the warm water, despite low difference between treatment and holding temperature (Δt = 13, 16 or 19 °C). Eye damages were more prevalent in the warm water treated groups than in the controls. Little difference was recorded between treatment groups in their growth and condition factor, blood plasma values, organ health, and long-term coping ability. There was, however, a significant increase in mortality as a function of temperature after the first treatment (14 °C: 6.5%, 27 °C: 5.3%, 30 °C: 12.4% and 33 °C: 18.9% mortality). The first treatment was performed only two weeks after the fish had been tagged and moved into the experimental holding tanks, while the fish had been allowed to recover for four weeks without any handling before the second treatment. The group of fish that were not subjected to any treatments (the negative control) had no mortality throughout the entire experimental period.

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.

Illuminating the planktonic stages of salmon lice: A unique fluorescence signal for rapid identification of a rare copepod in zooplankton assemblages.

Monitoring of planktonic salmon louse (Lepeophtheirus salmonis salmonis) abundance and parameterization of key life-history traits has been hindered by labour-intensive and error-prone quantification using traditional light microscopy. Fluorescence illumination has been proposed as a means of improving visualization, but prior to this study adequate investigation of the relevant fluorescence profiles and measurement conditions has not been undertaken. We investigated the fluorescence profiles of L. salmonis and non-target copepod spp. with excitation and emission matrices (200-600 nm) and identified unique fluorescence signals. Fluorescence microscopy using excitation wavelengths of 470 ± 40 nm, and emission wavelengths of 525 ± 50 nm, showed that after 90 days of formalin storage salmon lice have a mean fluorescence intensity that is 2.4 times greater than non-target copepods (copepodid and adult stages). A 7-day heat treatment of 42°C in formalin increased the difference between salmon louse copepodids and non-target copepods to a factor of 3.6, eliminating the need for prolonged storage. Differences in the fluorescence signal and endogenous fluorophores were investigated with respect to variation in sea lice species, age, stage and host fish origin. Under the conditions outlined in this paper, the fluorescence signal was found to be a reliable means of visualizing and differentiating salmon lice from non-target zooplankters. Adaptation of the fluorescence signal would greatly expedite traditional methods of enumerating salmon louse larvae in plankton samples and could provide a means of automated detection.

The effect of temperature on ability of Lepeophtheirus salmonis to infect and persist on Atlantic salmon.

The salmon louse (Lepeophtheirus salmonis) is an ecologically and economically important parasite of salmonid fish. Temperature is a strong influencer of biological processes in salmon lice, with development rate increased at higher temperatures. The successful attachment of lice onto a host is also predicted to be influenced by temperature; however, the correlation of temperature with parasite survival is unknown. This study describes the effects of temperature on infection success, and survival on the host during development to the adult stage. To accurately describe infection dynamics with varying temperatures, infection success was recorded on Atlantic salmon (Salmo salar) between 2 and 10°C. Infection success ranged from 20% to 50% and was strongly correlated with temperature, with the highest success at 10°C. Parasite loss was monitored during development at eight temperatures with high loss of lice at 3 and 24°C, whilst no loss was recorded in the temperature range from 6 to 21°C. Sea temperatures thus have large effects on the outcome of salmon louse infections and should be taken into account in the management and risk assessment of this parasite. Improving understanding of the infection dynamics of salmon lice will facilitate epidemiological modelling efforts and efficiency of pest management strategies.

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.

Salmon lice survive the straight shooter: A commercial scale sea cage trial of laser delousing.

Ectoparasitic salmon louse (Lepeophtheirus salmonis) infestations are costly for Atlantic salmon (Salmo salar) farmers in Norway. As a result, there is a strong desire for solutions to prevent and control infestations, and new technologies are typically developed and commercialised rapidly, without rigorous validation. Here, we tested the efficacy of a new commercially available control measure-delousing by underwater lasers-using a replicated design at full commercial scale. Laser delousing was used in combination with a preventive method (snorkel cages), with laser nodes deployed in 3 of the 6 sea cages at the site. The trial ran for 54 days, after which time there was no difference in infestation density of mobile salmon louse stages (pre-adult, adult male or adult female) in cages with or without laser nodes installed. By the end of the trial, adult female lice numbers in all cages were close to the legislated trigger for mandatory delousing (0.5 adult female lice per fish). The laser nodes delivered a large number of pulses relative to the number of lice in the cages, indicating that a lack of lethality rather than a lack of target detection was the limiting factor. If all pulses had been effective, they should have removed between 4-38 % of mobile lice each day. There was no effect on salmon welfare indicators such as skin condition or eye status. Our results highlight the importance of rigorous validation of new technologies across a range of conditions before widespread implementation by industry.

Where art thou louse? A snapshot of attachment location preferences in salmon lice on Atlantic salmon hosts in sea cages.

Problematic sea lice infestations on farmed Atlantic salmon (Salmo salar) have motivated extensive research and development into new methods to prevent, monitor and control sea lice. Most of these technologies require detailed information on the behaviour, spatial distribution and demography of lice on host fish. This study investigated how salmon lice (Lepeophtheirus salmonis) infestation density varies across the host's surface under sea cage farming conditions. Lice abundance, demography and attachment location were tracked over time, with repeated sampling of 300 individually tagged salmon across three replicate experimental sea cages. The data reveal clear differences in attachment locations according to sex and stage, but with an overall preference for the dorsal surface among mobile stages-dorsal head for adult females and dorsal-posterior section for males and pre-adults. Total lice abundance was highly variable between repeated measures of individual fish, consistent with frequent host-switching or mortality. Total lice numbers also declined between sampling dates, likely due to handling, with lost mobile lice being almost exclusively adult males. As the distribution of sea lice on hosts is likely determined by numerous factors, future image-based automated detection systems should be validated in settings that reflect the complex host-parasite interactions that occur in open farming systems.

Sterilization of sea lice eggs with ultraviolet C light: towards a new preventative technique for aquaculture.

BACKGROUND: Sea lice infestations on Atlantic salmon (Salmo salar) farms are a considerable burden on the industry and put wild salmonid populations at risk. Frequent delousing treatments are necessary to keep lice densities below allowable limits, but currently viable treatments have drawbacks in terms of financial cost, animal welfare, or environmental impacts. We tested if 254 nm ultraviolet C light (UVC) could function as a new preventative method to suppress reproduction of salmon lice (Lepeophtheirus salmonis) by sterilizing fertilized eggs. We exposed salmon lice eggstrings to a range of UVC intensities and durations to identify effective doses. RESULTS: A cumulative dose of 0.008 J cm-2 induced 5% egg mortality, while 95% egg mortality occurred at 0.09 J cm-2 , indicating that UVC can be effective as a preventative treatment. The total cumulative dose appeared to be more important than the duration or number of individual exposures by which the total dose was achieved. CONCLUSION: UVC treatment has immediate applications for the salmon aquaculture industry, including for the treatment of wastewater from delousing or other operations. Future work will assess the feasibility of UVC dose delivery on host salmon in sea cage environments that involves little or no fish handling and creates negligible environmental impacts. © 2019 Society of Chemical Industry.

To treat or not to treat: a quantitative review of the effect of biofouling and control methods in shellfish aquaculture to evaluate the necessity of removal.

The global growth of farmed shellfish production has resulted in considerable research investigating how biofouling compromises farm productivity. Shellfish fitness can be compared between fouled stock and stock which has undergone treatment. As treatment options are often harsh, they may deleteriously affect stock. The projected impact of biofouling may therefore be confounded by the impact of treatments. Given the substantial cost of fouling removal, some have questioned the necessity of biofouling mitigation strategies. Meta-analysis revealed that biofouling typically reduces shellfish fitness. However, the fitness of treated stock was often lower or equal to fouled control stock, indicating that many common antifouling (AF) strategies are ineffective at enhancing farm productivity. Overall, caution and diligence are required to successfully implement biofouling mitigation strategies. The need remains for increased passive prevention approaches and novel AF strategies suitable for shellfish culture, such as strategic siting of bivalve farms in areas of low biofouling larval supply.

Sea lice infestation levels decrease with deeper 'snorkel' barriers in Atlantic salmon sea-cages.

BACKGROUND: Salmon lice (Lepeophtheirus salmonis) are the most important parasites of farmed salmon. Infective larvae position themselves in the upper part of the water column to increase encounter probabilities with potential hosts. Previous studies have shown that a 'snorkel' sea-cage technology protects salmon from infection in surface waters. We tested whether deep snorkels would more effectively reduce lice infestation than shallow snorkels and still uphold adequate conditions for the fish. Five sea-cages (12 m × 12 m) each holding approximately 3000 Atlantic salmon (Salmo salar) (53 ± 10 g) were fitted with snorkels that gave protection from infection for 0, 4, 8, 12 or 16 m. We tested whether reductions in the settlement of new salmon lice copepodids were consistent among four separate infection periods. RESULTS: Lice infestation decreased exponentially with depth in all time periods. Infection levels in shallow snorkels (0 and 4 m) were consistently 4-10 times higher than those in deep snorkels (12 and 16 m). Key welfare and production performance indices were similar across all snorkel depths. CONCLUSION: Deeper snorkels dramatically and consistently reduced infection levels of salmon lice compared with shallow snorkels, without consequences for fish welfare and production performance. Therefore, reducing salmon sea lice encounters using a depth-based barrier is a powerful management tool for salmon farming. © 2017 Society of Chemical Industry.

Group behavioural responses of Atlantic salmon (Salmo salar L.) to light, infrasound and sound stimuli.

Understanding species-specific flight behaviours is essential in developing methods of guiding fish spatially, and requires knowledge on how groups of fish respond to aversive stimuli. By harnessing their natural behaviours, the use of physical manipulation or other potentially harmful procedures can be minimised. We examined the reactions of sea-caged groups of 50 salmon (1331 ± 364 g) to short-term exposure to visual or acoustic stimuli. In light experiments, fish were exposed to one of three intensities of blue LED light (high, medium and low) or no light (control). Sound experiments included exposure to infrasound (12 Hz), a surface disturbance event, the combination of infrasound and surface disturbance, or no stimuli. Groups that experienced light, infrasound, and the combination of infrasound and surface disturbance treatments, elicited a marked change in vertical distribution, where fish dived to the bottom of the sea-cage for the duration of the stimulus. Light treatments, but not sound, also reduced the total echo-signal strength (indicative of swim bladder volume) after exposure to light, compared to pre-stimulus levels. Groups in infrasound and combination treatments showed increased swimming activity during stimulus application, with swimming speeds tripled compared to that of controls. In all light and sound treatments, fish returned to their pre-stimulus swimming depths and speeds once exposure had ceased. This work establishes consistent, short-term avoidance responses to these stimuli, and provides a basis for methods to guide fish for aquaculture applications, or create avoidance barriers for conservation purposes. In doing so, we can achieve the manipulation of group position with minimal welfare impacts, to create more sustainable practices.