Safety and efficacy of emamectin benzoate administered in-feed to Atlantic salmon, Salmo salar L., smolts in freshwater, as a preventative treatment against infestations of sea lice, Lepeophtheirus salmonis (Krøyer)

The safety and efficacy of emamectin benzoate, administered in-feed to Atlantic salmon smolts, Salmo salar L., held in freshwater, was evaluated as a preventative treatment against sea lice, Lepeophtheirus salmonis, following transfer of fish to seawater.

In the safety study, salmon smolts held in freshwater were fed with diets containing emamectin benzoate at nominal doses of 0 (control), 50 (recommended dose) and 250 (5× recommended dose) μg kg⁻¹ fish day⁻¹ for 7 days (days 0–6). Actual dose rates, based on measured concentrations of emamectin benzoate in feed, differences in fish weight, and feed consumed, were 0, 54, and 272 μg kg⁻¹ day⁻¹, respectively. On day 9, fish were transferred to seawater and observed for 14 days. No differences in feeding response, coordination, behaviour, gross and histological appearance were observed between control fish and those that received 54 μg kg⁻¹ day⁻¹. Among smolts that received 272 μg kg⁻¹ day⁻¹, approximately 50% exhibited darker coloration, and one fish (1%) exhibited uncoordinated swimming behaviour. No pathognomonic signs of emamectin benzoate toxicity were identified.

In the efficacy study, smolts held in freshwater were fed an unmedicated ration (control group) or emamectin benzoate at 50 μg kg⁻¹ day⁻¹ (treated group) for 7 days (days 0–6). On day 9, fish were re-distributed to eight seawater tanks, each holding 30 control and 30 treated fish. On days 28, 56, 77 and 109, respectively, control and treated fish in two tanks were challenged with L. salmonis copepodites. When lice in each group reached chalimus stage IV, fish were sampled and the numbers of lice were recorded. Fish challenged at day 109 were sampled for the second time when lice were at the adult stage. Efficacy was calculated as the reduction in the mean number of lice on treated fish relative to the mean on control fish. Treatment with emamectin benzoate resulted in an efficacy of 85.0–99.8% in fish challenged at days 28–77, from the start of treatment, and lice counts were significantly lower (P<0.001) on treated fish than on controls. When fish challenged at day 109 were sampled at day 128, efficacy was 44.3%, but survival of chalimus to adult lice on treated fish was lower, and at day 159, efficacy had increased to 73%. These results demonstrate that treatment of salmon smolts with emamectin benzoate in freshwater was well tolerated and highly effective in preventing sea lice infestation following transfer of fish to seawater.     

The efficacy of emamectin benzoate as an oral treatment of sea lice, Lepeophtheirus salmonis (KrÒyer), infestations in Atlantic salmon, Salmo salar L.

The efficacy of emamectin benzoate as an oral treatment of sea lice, Lepeophtheirus salmonis (KrÒyer), infestations in Atlantic salmon, Salmo salar L., was evaluated in a dose titration study and two dose confirmation studies. Replicated groups of salmon with induced infestations of sea lice were given emamectin benzoate on pelleted feed at doses of 0, 25, 50 and 100 μg kg⁻¹ biomass day⁻¹ for seven consecutive days. Sea lice were counted at 7, 14 and 21 days from the start of treatment, and comparisons made with control fish given the same diet without emamectin benzoate. Total numbers of sea lice were significantly reduced at all doses in all three studies when compared to control fish. There was no significant difference between doses of 50 and 100 μg kg⁻¹, but the 50 μg kg⁻¹ dose resulted in significantly fewer lice than the 25 μg kg⁻¹ dose. Emamectin benzoate was highly effective in reducing numbers of preadult and adult lice and prevented the maturation of chalimus to motile stages. The optimum therapeutic dose was selected as 50 μg kg⁻¹ day⁻¹ for seven days. Treatment reduced the incidence of epidermal damage by sea lice and, in one study, survival of treated fish was 48% higher than in control groups. No fish mortalities or adverse effects were attributed to treatment with emamectin benzoate at any of the doses tested.     

Efficacy of teflubenzuron (Calicide®) for the treatment of sea lice, Lepeophtheirus salmonis (Krøyer 1838), infestations of farmed Atlantic salmon (Salmo salar L.)

The efficacy of teflubenzuron (Calicide^®) for the treatment of farmed Atlantic salmon (Salmo salar L.) infested with sea lice [Lepeophtheirus salmonis (Krøyer)] was investigated on two commercial salmon farms. Teflubenzuron (Calicide^®) coated onto commercial feed pellets was administered orally at 10 mg per kg body weight per day for seven consecutive days. Fish were randomly sampled and lice numbers recorded from both treated and control groups on four sampling occasions post‐medication. Statistically significant reductions in the number of L. salmonis per fish were recorded. Maximum efficacy was observed toward chalimus and preadult stages of L. salmonis, and was achieved about 7 days post‐medication. No apparent effect was observed on adult lice. No adverse drug reactions or palatability problems were associated with the treatments.     

Interspecific comparison of experimental and natural routes of Lepeophtheirus salmonis and Caligus elongatus challenge and consequences for distribution of chalimus on salmonids and therapeutant screening

Although the gills have been reported as a site for settlement by sea lice attached stages in experimental challenges, the prevalence and abundance of infection on gills have not been reported in farmed fish and have been reported only rarely in wild salmonids. Disproportionate attachment to the gills may be significant in evaluation of new therapeutants and this study asks whether the efficacy data from studies of therapeutants based on experimental challenge can be extrapolated to the fish farm arena. The distribution of chalimus on gills, body and fins of farmed first sea year Atlantic salmon were examined in two Scottish sea lochs, each on two occasions (30 fish on each sampling date), fish from one farm with infection with Lepeophtheirus salmonis only and the other farm with exclusively Caligus elongatus. The dorsal fin was the primary attachment site of L. salmonis (50% and 33% in two samples respectively) with 29% and 32% on other fins and 20% and 29% on the body and head. Only one of 56 chalimus (1.8%) and four of 75 chalimus (5.3%) were attached to the gills. In contrast, the chalimus distribution of C. elongatus was predominantly on fins other than the dorsal, including the tail, 74% and 71%, on two sampling occasions, respectively, with only 4.2% and 3.6% of chalimus located on the dorsal fin. The pectoral fins were the most common location with 32%, followed by the tail 23%, pelvic fins 14% and only 3% on the anal fin. None of 287 and 111 chalimus in two samples, respectively, of C. elongatus was attached to the gills. The low abundance of lice on the gills indicated that non‐destructive sampling methods adequately represented lice infection. In three experimental challenges with L. salmonis, the gills were a major attachment location with 19.9–36.3% of chalimus, 9.9–26.9% on the body, 22–27% on the dorsal fin and 19–23% on the other fins. These differences with natural challenge in L. salmonis reported here and also in the literature suggest that the importance of the gills for chalimus location in the experimental model is atypical and an artefact of reduced host swimming speed in tanks that permits copepodids to attach to the gills. In natural infection, the flow over the gills is high and reduced flow is required for copepodid settlement. Greatest reductions (96%) in chalimus bath treated with cypermethrin occurred on the gills with no significant reductions in those located on the dorsal and other fins. It is possible that the efficacy of therapeutants may be higher in chalimus attached to the gills because of the higher concentration encountered of bath therapeutant and extrapolation of efficacy to farms may be misleading and progression to field testing is essential.     

An evaluation of the duration of efficacy of emamectin benzoate in the control of Caligus curtus Müller infestations in Atlantic cod, Gadus morhua L

The duration of efficacy of emamectin benzoate in the control of Caligus curtus infestations in Atlantic cod, Gadus morhua L., was studied following an administration of 50 μg kg^?1 for seven consecutive days. No lice were found on medicated fish when challenged 1 week (challenge 1) or 5 weeks (challenge 2) following termination of medication, whereas the mean abundance of lice among the unmedicated fish was 17.9 and 19.3 lice per fish in challenge 1 and 2, respectively. Muscle concentrations of 19.5 ± 8.2 ng g^?1 and 3.4 ± 0.9 ng g^?1, respectively, and skin concentrations of 23.1 ± 10.8 and 4.2 ± 1.0 ng g^?1, respectively, were found 27 and 55 days following the termination of medication. Tissue concentrations and the duration of efficacy indicate a dosing regime for emamectin in cod, similar to the regime used for Atlantic salmon, Salmo salar L.     

The settlement and reproductive success of Lepeophtheirus salmonis (Krøyer 1837; Copepoda: Caligidae) on atypical hosts

The reproductive success of Lepeophtheirus salmonis settled on host and non‐host fish has been compared. Triplicate single species tanks of Atlantic salmon, marine three‐spined sticklebacks, saithe and Atlantic cod were exposed to 10 adult female L. salmonis per tank (n=30 lice per species). Adult female L. salmonis settlement and egg string production occurred only on salmon and cod, with no egg production occurring on saithe and three‐spined sticklebacks. The number of eggs in egg strings, hatching success of eggs and the survival of all larval stages to the copepodid stage were severely affected by the species of fish on which female L. salmonis had settled. L. salmonis settled on cod produced significantly fewer eggs, lower hatching rates and lower survival rates of larvae than females on Atlantic salmon. The production of egg strings by L. salmonis females infecting cod, which successfully hatch and moult through to the infective copepodid stage, albeit in small numbers, is discussed in terms of the implications to aquaculture and salmon and cod farming scenarios.     

Laboratory and field investigations of salmon lice [Lepeophtheirus salmonis (Krøyer)] infestation on Atlantic salmon (Salmo salar L.) post-smolts

Hatchery‐reared 1‐year‐old Atlantic salmon post‐smolts (Salmo salar L.), artificially infected with salmon lice [Lepeophtheirus salmonis (Krøyer)] copepodids, were found to suffer from primary alterations (increased cortisol levels) at early lice stages. Secondary alterations, such as osmotic stress (increased chloride levels), first occurred after the preadult stages of the lice appeared. Fish with the highest salmon lice infections died throughout the experiment. Seven years of field investigation of Trondheimsfjorden showed that Atlantic salmon post‐smolts descending coastal waters can become heavily infected with salmon lice. The migrating post‐smolts were only infected with the chalimus stages, showing that the fish had only recently left the rivers. The infection level, however, varied considerably between the years, and, in 1998, the infection was higher than previous years. The experimental results have been combined with the field data to appraise the consequences of the infection.     

Further evaluation of the efficacy of emamectin benzoate for treating Pseudocapillaria tomentosa (Dujardin 1843) in zebrafish Danio rerio (Hamilton 1822)

Pseudocapillaria tomentosa is a pathogenic nematode parasite, causing emaciation and severe inflammatory lesions in the intestines in zebrafish Danio rerio (Hamilton 1822). Emamectin benzoate is commercially available analogue of ivermectin used for treating salmon for sea lice, under the brand name SLICE^®, and we have used this for treating zebrafish with the P. tomentosa. Here, SLICE^®, 0.2 per cent active emamectin benzoate, was used for oral treatments at 0.35 mg emamectin benzoate/kg fish/day for 14 days starting at 7 days post‐exposure (dpe). Another experiment entailed initiating treatment during clinical disease (starting at 28 dpe). Early treatment was very effective, but delaying treatment was less so, presumably due to inappetence in clinically affected fish. We evaluated emamectin benzoate delivered in water, using Lice‐Solve? (mectinsol; 1.4% active emamectin benzoate) in two experiments. Application of four 24‐hr treatments, space over 7 days was initiated at 28 dpe at either 0.168 or 0.56 mg emamectin benzoate/L/bath, and both treatments completely eradicated infections. This was 3 or 10 times manufacture's recommended dose, but was not associated with clinical or histological side effects.     

Evaluation of sea lice abundance levels on farmed Atlantic salmon (Salmo salar L.) located in the Broughton Archipelago of British Columbia from 2003 to 2005

Salmon farming began in British Columbia (BC) in the 1970s and in 2006, aquaculture represented BC's largest agricultural export. Along with this growth in production has been a growth in controversy, including the concern that sea lice originating from Atlantic salmon farms negatively impact wild juvenile pink salmon in the Broughton Archipelago. To understand the dynamic interaction between wild and farmed fish, data for on‐farm abundance of sea lice are required. In this study, 33 000 Atlantic salmon from 20 active farms were examined over 3 years. Two species of lice were found: Lepeophtheirus salmonis and Caligus clemensi. Inter‐annual and seasonal variations in abundance levels occurred with lower levels of L. salmonis in 2003 compared with 2004 and 2005, while C. clemensi levels were highest in 2003. The abundance of L. salmonis was greater on older farmed fish. The findings are compared with European and eastern Canadian sea lice reports, and possible sources of sea lice on farmed salmon are discussed.     

A comparison of epidemiological patterns of salmon lice, Lepeophtheirus salmonis, infections on farmed Atlantic salmon, Salmo salar L., in Norway and Scotland

This paper examines two large national data sets collected over several years and contrasts the patterns of sea lice, Lepeophtheirus salmonis (Krøyer, 1837), infections, the use of treatments and the occurrence of chalimus peaks between Norwegian and Scottish farms. Infection levels in Scotland were significantly higher in general over the period under study. For the chalimus stage group in the first quarter of the year, Norwegian mean abundance stayed below 10 lice per m² while Scottish means reached 45 lice per m² of fish skin per m³ of water. Both countries had more chalimus in summer than at other times of year, but in the last 3 months of the year Scottish fish had, on average, two to four times as many chalimus as Norwegian fish. Peaks of chalimus abundance were more frequent in Scotland, particularly in winter, but the most prominent peaks occurred in summer in both countries. In Scotland a marked mid-year build-up of mobile pre-adult and adult stages was seen, and both countries showed a tendency for mobile counts on the second year fish to increase towards the end of the year. Scottish fish carried, on average, three times as many mobile lice per m² of skin as Norwegian fish in the last 3 months of the year. The difference in lice loads was reflected in the greater use of veterinary medicines on Scottish farms. The higher infection levels in Scotland may be due to shallower and more enclosed water bodies used for farming, smaller and shallower pens, differences in sea water temperatures or in access to appropriate medication. The results highlight the importance of ensuring that effective veterinary medicines are available in the UK for the control of infection.     

Does the AquasmartTM feeding system reduce sea louse [Lepeophtheirus salmonis (Krøyer)] infestation on farmed Atlantic salmon (Salmo salar L.) in winter?

Infections of sea lice [Lepeophtheirus salmonis (Krøyer)] were studied over winter on a working, commercial salmon farm in relation to two types of feeding system: a regular, pneumatic spray feeder and an irregular, ‘demand feed’ sensor system (Aquasmart^TM). Contrary to prediction, the Aquasmart^TM system alone had no significant effect on chalimus infestation compared with fish fed by pneumatic spray. However, it was associated with significantly reduced numbers of mobile stages, both at the beginning of the study (P < 0.05) and, more particularly, in conjunction with treatment of all pens using hydrogen peroxide (P < 0.001). It is suggested that feeding regime may represent a useful tool in reducing lice infestation when co‐ordinated with other methods of control, such as peroxide treatment, and that further work in this field is merited.     

Early‐stage sea lice recruits on Atlantic salmon are freshwater sensitive

Sea lice are significant parasites of marine and brackish farmed fishes. Freshwater bathing is a potential control option against numerous sea lice species, although has been viewed as futile against those that are capable of tolerating freshwater for extended periods. By comparing freshwater survival times across host‐attached stages of Lepeophtheirus salmonis (Krøyer), a key parasite in Atlantic salmon farming, we show the first attached (copepodid) stage undergoes 96–100% mortality after 1 h in freshwater, whereas later attached stages can tolerate up to 8 days. Thus, regular freshwater bathing methods targeting the more susceptible attached copepodid stage may successfully treat against L. salmonis and potentially other sea lice on fish cultured in marine and brackish waters.     

How are the salmon lice (Lepeophtheirus salmonis Krøyer, 1837) in Atlantic salmon farming affected by different control efforts: A case study of an intensive production area with coordinated production cycles and changing delousing practices in 2013–2018

The aims of the present study were to describe the salmon lice (Lepeophtheirus salmonis Krøyer, 1837) situation in an intensive salmon production area in mid‐Norway and to consider implications of changing practices of how salmon lice infestation can be controlled. The results in this study suggest that there are steps that can be carried out to keep salmon lice under control even during years when the temperature facilitates a quick salmon lice development. The present work indicates that the use of cleaner fish can delay the time it takes adult female lice to reach 0.1 per salmon in the beginning of a production cycle. It suggests that the timing of cleaner fish deployment into salmon cages can influence its effectiveness in controlling salmon lice. It also gives caution to letting salmon lice develop unchecked, even at levels far below the current lice limit, because of the difficulties to control salmon lice when the external infection pressure is too high. This study took place during a rapid change in delousing methods, in an area with coordinated salmon production. Despite its exploratory nature, this study offers insights into the salmon lice fluctuations in relation to efforts aimed at controlling it.     

The Three‐spined Stickleback,Gasterosteus aculeatus Linnaeus 1758, plays a minor role as a host of Lepeophtheirus salmonis (Krøyer 1837) in the Gulf of Maine

The sea louse, Lepeophtheirus salmonis (Krøyer 1837), is a significant parasite of farmed salmon throughout the Northern Hemisphere. Management of on‐farm louse populations can be improved by understanding the role that wild fish play in sustaining and providing refuge for the local population of sea lice. In this study, 1,064 sticklebacks were captured. Of these animals, 176 individuals were carrying a total of 238 sea lice, yielding a prevalence and intensity of 16.5% and 1.4 lice per fish, respectively. Detailed examination of the sea lice on the three‐spined sticklebacks captured in Cobscook Bay found two L. salmonis individuals using three‐spined sticklebacks as hosts. A 2012 survey of wild fish in Cobscook Bay, Maine, found multiple wild hosts for Caligus elongatus (von Nordmann 1832), including three‐spined sticklebacks (Gasterosteus aculeatus L.), but no L. salmonis were found in this earlier study.     

Survival of salmon lice, Lepeophtheirus salmonis Krøyer, on Arctic charr, Salvelinus alpinus (L.), in fresh water

The survival of salmon lice. Lepeophtheirus salmonis (Krøyer) on Arctic charr. Salvelinus alpinus (L.). in fresh water was studied. The results showed that salmon lice were able to survive for up to 3 weeks, in contrast with previous published information that the parasite is quickly shed when the fish enters fresh water.     

Efficacy of emamectin benzoate against sea lice infestations of Atlantic salmon, Salmo salar L.: evaluation in the absence of an untreated contemporary control

The efficacy of emamectin benzoate (SLICE) against sea lice infestations of Atlantic salmon, Salmo salar L., is typically assessed using untreated fish, or fish treated with alternative therapeutants, as controls. The State of Maine, USA, is currently under active management for the OIE-notifiable pathogen, infectious salmon anaemia virus (ISAV); consequently, neither control group is feasible in this region. Untreated salmon risk extensive damage from the ectoparasites, and threaten to increase vector-borne exposure or susceptibility of farms to ISAV; and the only treatment presently available in Maine is SLICE. However, because sea lice infestations are unlikely to resolve spontaneously, and response to treatment occurs within weeks, use of a pretreatment baseline is a reasonable alternative for confirmatory studies. We evaluated SLICE efficacy on Atlantic salmon farms in Cobscook Bay 2002-2005, in the absence of untreated controls, using pretreatment lice loads as a reference for calculation. Maximum efficacy ranged from 68% to 100% reduction from initial levels. Time-to-maximum efficacy ranged from 1 to 8 weeks after treatment initiation. Efficacy duration, measured between first reduction and first progressive rise in counts, ranged from 4 to 16 weeks.     

Differences in Lepeophtheirus salmonis abundance levels on Atlantic salmon farms in the Broughton Archipelago, British Columbia, Canada

Sea lice data collected from Atlantic salmon farms in the Broughton Archipelago between 2003 and 2005 were examined for inter-regional differences in mobile Lepeophtheirus salmonis (Krøyer) abundance using the generalized linear model procedure. Factors such as age of the salmon populations, location of farms and time of year had a significant effect on the abundance of the mobile stages of L. salmonis whereas water temperature and salinity did not. Separate evaluation of SLICE® treatment data found no significant difference in treatment frequency among the areas but did show that there were significantly lower numbers of farm treatments during the summer months when compared with other seasons. The role of environment and wild fish in influencing sea lice abundance on the farmed salmon is discussed. The findings suggest that effective management programmes for sea lice should not only be based on geographical location but should take into account other factors which could influence lice abundance levels.     

Environmental effects of the anti-sea lice (Copepoda: Caligidae) therapeutant emamectin benzoate under commercial use conditions in the marine environment

Sea lice infestation can pose a particular problem to intensive cage aquaculture of salmonids. It is most often treated by therapeutants added to the water or incorporated into fish feed. This study investigated the environmental effects of one such in-feed treatment, emamectin benzoate, under commercial use conditions. Atlantic salmon were fed the commercial preparation of emamectin benzoate at a production fish farm on the west coast of Scotland. Sediment dwelling fauna, large mobile fauna and sentinel organisms were monitored for potential impacts over a 1 year post-treatment period (23 September 1997 to 23 September 1998). Additionally, sediment and water samples, transplanted blue mussels and captured large fauna were analysed for accumulated emamectin benzoate and its desmethylamino metabolite to investigate the long term environmental fate of emamectin benzoate.

Results from analysis of sediments suggested that limited deposition of emamectin benzoate took place during and up to 1 week post-treatment, and may have continued up to 4 months of post-treatment. This was consistent with the release of the active ingredient from fish excretion and defecation after treatment. Quantifiable amounts of emamectin benzoate were found 10 m from the cages 12 months after treatment. However, clear reduction in overall sediment concentrations indicated that the deposited emamectin was degrading over time. Emamectin benzoate was not detected in water samples during the field trial. Quantifiable concentrations of emamectin benzoate were found in blue mussels deployed up to 100 m from the treatment cages at 1 week post treatment, but found only at 10 m from the cages at 1 month post-treatment. This suggests that while mussels were actively accumulating emamectin benzoate immediately after treatment by 1 month this had largely been depurated.

Macrobenthic faunal analysis indicated that there was no evidence that the occurrence of emamectin benzoate, or its desmethylamino metabolite, in sediments around fish farm cages after treatment had any toxic impacts on organisms in either water column or sediments.     

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.     

Resistance of sea lice, Lepeophtheirus salmonis (Krøyer), to hydrogen peroxide on farmed Atlantic salmon, Salmo salar L.

Hydrogen peroxide has been the only medicine used to treat salmon infected with sea lice [Lepeophtheirus salmonis (Krøyer)] on many farms in Scotland since 1992 and reports have suggested reduced treatment efficacy. The present study tests the sensitivity of sea lice to exposure to hydrogen peroxide under farm conditions and also in vivo by comparing lice from a farm with suspected resistance and lice from a farm that had not been pre‐exposed to hydrogen peroxide. In bin treatments on fish from a farm treated with hydrogen peroxide on 41 occasions, numbers of ovigerous lice declined by only 15% and 16% in two replicates and other mobile stages by 25%. Where hydrogen peroxide had not been used, ovigerous females declined by 90% and 87% and other mobile lice stages by 97% and 99%. These trials and observed poor efficacy of cage treatments, for example only 7.5% reduction in lice numbers when fish were treated with 2000 p.p.m. for 20 mins, indicated resistance of lice to hydrogen peroxide. The mechanisms involved in the development of resistance, possible genetic selection for lice with reduced carapace permeability or detoxifying enzymes such as catalase, or tolerance through induction by subtherapeutic exposure are reviewed. Implications for lice control strategies relying on hydrogen peroxide are discussed.