Effects of the sea lice chemotherapeutant, emamectin benzoate, on metabolism and behaviour of the sea-pen Pennatula phosphorea.

Chemotherapeutants used to control infestations by sea lice can be released into the marine environment surrounding aquaculture farms. Among these therapeutic agents, emamectin benzoate is extensively utilized even though its impact on non-target taxa has not been thoroughly examined. In this context, we explored the effects of emamectin benzoate on a common Norwegian habitat-forming species: the phosphorescent sea-pen Pennatula phosphorea. Specifically, we examined P. phosphorea metabolic and responses before, during and after exposure to emamectin benzoate. Results indicate that an 8-day emamectin benzoate exposure (0.8 mg/L) did not induce P. phosphorea mortality or significant behavioural or metabolic modifications. However, we highlighted the presence and persistence of emamectin benzoate in exposed P. phosphorea tissue. These results indicate that emamectin benzoate is unlikely to adversely impact P. phosphorea populations in the environment. However, persistence of emamectin benzoate in tissue constitutes a potential for bioaccumulation with repeated treatments and should be examined in further studies.

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.

CO2-Induced Ocean Acidification Alters the Burrowing Behavior of Manila Clam Ruditapes philippinarum by Reversing GABAA Receptor Function.

Biological burrowing behavior is an important driver shaping ecosystems that is being threatened by CO2-induced ocean acidification; however, the effects of ocean acidification on burrowing behavior and its neurological mechanism remain unclear. This study showed that elevated pCO2 significantly affected the burrowing behaviors of the Manila clam Ruditapes philippinarum, such as increased foot contraction, burrowing time, and intrabottom movement and decreased burrowing depth. Delving deeper into the mechanism, exposure to elevated pCO2 significantly decreased extracellular pH and increased [HCO3-]. Moreover, an indicator GABAA receptor, a neuroinhibitor for movement, was found to be closely associated with behavioral changes. In situ hybridization confirmed that the GABAA receptor was widely distributed in ganglia and foot muscles, and elevated pCO2 significantly increased the mRNA level and GABA concentration. However, the increase in GABAA receptor and its ligand did not suppress the foot movement, but rather sent "excitatory" signals for foot contraction. The destabilization of acid-base homeostasis was demonstrated to induce an increase in the reversal potential for GABAA receptor and an alteration in GABAA receptor function under elevated pCO2. This study revealed that elevated pCO2 affects the burrowing behavior of Manila clams by altering GABAA receptor function from inhibitory to excitatory.

Effect of sea lice chemotherapeutant hydrogen peroxide on the photosynthetic characteristics and bleaching of the coralline alga Lithothamnion soriferum.

The proliferation of sea lice (Lepeophtheirus salmonis) represents a major challenge for the salmonid aquaculture industry in Norway. Hydrogen peroxide (H2O2) is a chemotherapeutant frequently used on Norwegian farms, however, its toxicity to non-target benthic species and habitats remains poorly understood. Maerl beds are constructed by the accumulation of non-geniculate coralline algae and provide important ecological functions. Due to the rapid expansion of aquaculture in Norway and the continued use of H2O2 as an anti-sea lice treatment, it is crucial to understand the impact of H2O2 on the physiology of maerl-forming species. The effects of a 1 h exposure to H2O2 on the photophysiology and bleaching of the coralline alga Lithothamnion soriferum were examined here through a controlled time-course experiment. PAM fluorimetry measurements showed that H2O2 concentrations ≥ 200 mg l-1 negatively affected photosystem II (PSII) in thalli immediately after exposure, which was observed through a significant decline in maximum photochemical efficiency (Fv/Fm) and relative electron transport rate (rETR). The negative effects on PSII induced by oxidative stress, however, appear to be reversible, and full recovery of photosynthetic characteristics was observed 48 h to 28 days after exposure to 200 mg H2O2 l-1 and 2000 mg H2O2 l-1, respectively. At 28 days after exposure, there was evidence of two- to four-times more bleaching in thalli treated with concentrations ≥ 200 mg H2O2 l-1 compared to those in the control. This indicates that despite the recovery of PSII, persistent damages can occur on the structural integrity of thalli, which may considerably increase the vulnerability of coralline algae to further exposure to H2O2 and other chemical effluents from salmonid farms.

Antibacterial treatment of lumpfish (Cyclopterus lumpus) experimentally challenged with Vibrio anguillarum, atypical Aeromonas salmonicida and Pasteurella atlantica.

Lumpfish is a novel farmed species used as cleaner fish for the removal of lice from farmed salmon. As often with new, farmed species, there are challenges with bacterial infections. The frequency of prescription of antibiotic agents to lumpfish is increasing, despite the lack of knowledge about appropriate doses, duration of treatment and application protocols for the various antibacterial agents. In the current study, we have tested the effect of medicated feed with florfenicol (FFC), oxolinic acid (OA) and flumequine (FLU) on lumpfish experimentally challenged with Vibrio anguillarum, atypical Aeromonas salmonicida and Pasteurella atlantica. We found that all three antibacterial agents efficiently treated lumpfish with vibriosis using 10 and 20 mg kg-1  day-1 of FFC, 25 mg kg-1  day-1 of OA and 25 mg kg-1  day-1 FLU, whereas only FFC (20 mg kg-1  day-1 ) had good effect on lumpfish with pasteurellosis. None of the antibacterial agents were efficient to treat lumpfish with atypical furunculosis. FFC 20 mg kg-1  day-1 showed promising results in the beginning of the experiment, but the mortality increased rapidly 14 days post-medication. Efficient treatment is important for the welfare of lumpfish and for reducing the risk of development of antibiotic-resistant bacteria. To our knowledge, this is the first study to establish protocols for antibacterial treatment of lumpfish.

The Acute and Delayed Mortality of the Northern Krill (Meganyctiphanes norvegica) When Exposed to Hydrogen Peroxide.

Bath treatment pharmaceuticals used to control sea lice infestations in the salmonid industry, such as hydrogen peroxide (H2O2), are released directly into the environment where non-target organisms are at risk of exposure. The aim of this study was to determine the threshold concentrations for mortality of the Northern krill, Meganyctiphanes norvegica, a major component of the north Atlantic marine ecosystem. To assess the lethal effects of H2O2, we carried out a series of 1 h acute toxicity tests and assessed mortality through a 48 h post-exposure period. One-hour exposure to 170 mg/L, corresponding to 10% of the recommended H2O2 treatment, caused 100% mortality and a subsequent acute median-lethal concentration LC50 value of 32.5 mg/L. Increased mortality was observed with time in all exposed groups, resulting in successively lower LC50 values during the post-exposure period. The suggested H2O2 concentrations have the potential of causing negative effects to the Northern krill.

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.

Short-term exposure to hydrogen peroxide induces mortality and alters exploratory behaviour of European lobster (Homarus gammarus).

Bath treatment chemotherapeutants, used to control sea lice infestations in the salmonid aquaculture industry, are released directly into the marine environment around fish farms and pose a serious risk to non-target species, particularly crustaceans. Hydrogen peroxide (H2O2) is the most frequently used bath treatment chemotherapeutant on Norwegian fish farms, however, limited information is available on its toxicity to European lobsters (Homarus gammarus), a commercially important species at risk of exposure due to its distribution overlapping with salmon farm locations. The aim of this study was to investigate the lethal effects of H2O2 on pelagic (stage I-IV) larvae/post-larvae and its sub-lethal effects on the benthic stage V H. gammarus. To assess the lethal effects of H2O2, we carried out a series of 1 h toxicity tests and assessed mortality after a 24 h post-exposure period. Exposure to H2O2 was toxic to all pelagic larval stages tested, with estimated median lethal concentrations (LC50) of 177, 404, 665 and 737 mg/L for stage I, II, III and IV, respectively. These concentrations represent approximately 10, 23, 40 and 43%, of the recommended H2O2 concentrations used for delousing salmon on Norwegian fish farms, respectively. To assess the sub-lethal effects of H2O2 on H. gammarus, stage V juveniles were exposed to H2O2 at concentrations of 85, 170 and 510 mg/L for 1 h and shelter-seeking behaviour and mobility endpoints were assessed. Numerous behavioural parameters including distance travelled to shelter, time to locate shelter and the number of shelter inspections, were negatively affected in lobsters exposed to H2O2 when assessed immediately after the exposure period. However, no differences between control and exposed lobsters were detected after a 24 h post-exposure period. Our results demonstrate that short term exposures to H2O2 are lethal to pelagic H. gammarus life stages and can negatively affect the shelter seeking behaviour of benthic life stages, though these behavioural changes may be short-lived.

The impact of anti-sea lice pesticides, azamethiphos and deltamethrin, on European lobster (Homarus gammarus) larvae in the Norwegian marine environment.

Anti-sea lice pesticides, used in the salmonid aquaculture industry, are a growing environmental concern due to their potential to adversely affect non-target crustaceans. Azamethiphos and deltamethrin are two bath treatment pesticides used on salmon farms in Norway, however, limited information is available on their impact on European lobster (Homarus gammarus) larvae in the Norwegian marine environment. Here, we firstly report the lethal (LC50) and effective (EC50) concentrations of azamethiphos and deltamethrin for stage I and stage II larvae, following 1-h exposures. Using a hydrodynamic model, we also modelled the dispersal of both compounds into the marine environment around selected Norwegian farms and mapped the potential impact zones (areas that experience LC50 and EC50 concentrations) around each farm. Our data shows that azamethiphos and deltamethrin are acutely toxic to both larval stages, with LC50 and EC50 values below the recommended treatment concentrations. We also show that the azamethiphos impact zones around farms were relatively small (mean area of 0.04-0.2 km2), however deltamethrin impact zones covered much larger areas (mean area of 21.1-39.0 km2). These findings suggest that deltamethrin poses a significant risk to European lobster in the Norwegian marine environment while the impact of azamethiphos may be less severe.

Pharmacokinetic Data Show That Oxolinic Acid and Flumequine Are Absorbed and Excreted Rapidly From Plasma and Tissues of Lumpfish.

This study examined the uptake, tissue distribution and elimination of the antibacterial agents oxolinic acid and flumequine in lumpfish (Cyclopterus lumpus L.) by use of LC-MS/MS following a single oral administration of 25 mg/kg fish given in feed. Lumpfish are increasingly used as cleaner fish for removal of sea lice on commercially farmed salmon. The production of lumpfish is successful, but there are challenges with bacterial infections and the number of antibacterial treatments has increased in recent years. As the lumpfish is a novel species to farming, there is a need for pharmacokinetic data and establishment of protocols for efficient antibacterial treatment. The current study describes the pharmacokinetic properties of oxolinic acid and flumequine in lumpfish. Absorption of oxolinic acid was moderate and was characterized by a calculated peak plasma concentration (Cmax) of 2.12 µg/ml after 10.3 h (Tmax) and an elimination half-life (t1/2ß) of 21 h. Area under curve (AUC) and AUC from 0 to 24 h (AUC0-24h) were calculated to be 60.9 and 34.0 h µg/ml, respectively. For flumequine, plasma Cmax was found to be 2.77 µg/ml after 7.7 h (Tmax) with t1/2ß of 22 h. The area under the curve (AUC) and AUC from 0 to 24 h (AUC0-24) were calculated as 104.3 and 50.3 h µg/ml, respectively. Corresponding Cmax values in muscle, liver, and head-kidney for oxolinic acid were 4.01, 3.04, and, 4.68 µg/g, respectively and Tmax of 11.1, 9.2, and 10.0 h, respectively. For flumequine, Cmax values of 4.16, 4.01, and 7.48 µg/g were obtained in muscle, liver, and head kidney, respectively, with corresponding Tmax values of 10.2, 10.3, and 6.0 h. Antimicrobial susceptibility values as determined by minimum inhibitory concentration (MIC) analyses against 28 isolates of Aeromonas salmonicida isolated from diseased lumpfish ranged from 0.06 to 15 µg/ml for oxolinic acid and 0.024 to 6.25 µg/ml for flumequine. Bimodal distributions in susceptibility to both oxolinic acid and flumequine were observed. The combination of pharmacokinetic properties and MIC data make possible calculation of efficient treatment doses, which are needed to improve the welfare of lumpfish and minimize development of antibiotic resistant bacteria.

Whole-animal accumulation, oxidative stress, transcriptomic and metabolomic responses in the pink shrimp (Pandalus montagui) exposed to teflubenzuron.

The benzoylurea chitin synthesis inhibitor teflubenzuron, widely used against sea lice in North Atlantic aquaculture, may pose an environmental threat to non-targeted crustaceans. In this experiment, laboratory acclimated pink shrimp (Pandalus montagui), a species found in fjords with Atlantic salmon farming, were exposed to dietary teflubenzuron for 46 days (control; low dose: 0.01 µg/g; high dose: 0.1 µg/g). The exposure doses represent 0.1% and 1% of a standard treatment dose for Atlantic salmon. Mortality and prevalence of deformities, pharmacokinetics, oxidative stress and transcriptomic and metabolomic profiling were used to assess the response to teflubenzuron exposure. Mortality in the high-dose group was 25% (five of 20 individuals). No control or low-dose group shrimps died. Phenotypic responses,i.e., leg deformities (0 control, 6 low, 8 high) and cloudy eyes (0 control, 3 low, 7 high), were observed in some surviving shrimps (control n = 15, low n = 17, high n = 15). Accumulated levels of teflubenzuron in shrimps from the high-dose group ranged from 4.7 to 369 ng/g wet weight. Transcriptomic profiling showed very few significantly altered genes in the exposed shrimps. Teflubenzuron-induced changes to the metabolome pointed to well-known effects of benzoylurea agents, with reduced levels of N-acetylglucosamine indicating an effect on chitin synthesis. The metabolomic profiling showed that teflubenzuron exposure was associated with reduced energy metabolism. Some metabolites pointed to increased necrosis and/or bacterial overgrowth in the teflubenzuron-exposed shrimps. In conclusion, this study shows that teflubenzuron causes phenotypic effects in P. montagui exposed to 0.1% of the treatment dose given to Atlantic salmon.

Exposure to teflubenzuron negatively impacts exploratory behavior, learning and activity of juvenile European lobster (Homarus gammarus).

Infestations with salmon lice, a parasitic copepod, is a major problem in the salmon farming industry. Teflubenzuron is an in-feed pharmaceutical applied to control lice outbreaks; the standard medication is 10 mg per kg fish per day for seven days. Surveys reveal that teflubenzuron accumulates and persists in the sediment around fish farms and causes deformities and mortality in juvenile European lobster (Homarus gammarus), a species commonly found in the vicinity of salmon farms in Norway. To date, there is no information on sub-lethal effects of teflubenzuron on, for example, behavior. We conducted an experiment to assess possible difference in the shelter seeking behavior of teflubenzuron-exposed (N = 19) vs. not exposed (N = 19) H. gammarus juveniles. The teflubenzuron-exposed juveniles had been given very low concentrations, 1.7 µg per pellet twice per week for 113 days prior to this experiment. The concentration of teflubenzuron was estimated to be less than 1 ng/g lobster when they were tested in the behavior experiment. Animals were placed in a lane with a shelter at one end. Once a lobster had found and entered the shelter, they were repeatedly displaced back to the opposite end of the lane, for a total of 3 repeated runs per animal. Three of the exposed juveniles failed to settle in the shelter, and the remaining teflubenzuron-exposed animals took significantly more time to explore the environment and to find and recognize shelter. Furthermore, exposed lobsters also exhibited slower walking speed compared to the controls. These results demonstrate that teflubenzuron significantly reduces exploratory behavior, learning and activity of juvenile H. gammarus. Thus, exposure to teflubenzuron could increase predation mortality of juvenile lobsters in the wild.

Impact of teflubenzuron on the rockpool shrimp (Palaemon elegans).

Concerns have been raised over the environmental impacts of antiparasitic drugs used to delouse farmed salmon. Released into the marine environment, some of these drugs can have negative impact on non-targeted crustaceans in the vicinity of farming facilities. In this study, we examined the molecular effect of the insecticide teflubenzuron on a shrimp species inhabiting the littoral zone, the rockpool shrimp (Palaemon elegans). Rockpool shrimp was exposed for 98days to a dose representing 2% of a regular teflubenzuron medication applied to Atlantic salmon. Accumulation of teflubenzuron was studied in whole body samples, except abdominal segments 5 and 6, which were used for gene expression analysis. Insight into sublethal mode of action was sought by examining the transcriptional responses of 38 genes encoding proteins linked to molting and exoskeleton change, stress and detoxification. The accumulated levels of teflubenzuron in exposed animals varied between 1.7 and 33.0ng/g. Significant transcriptional effects of exposure were seen for markers linked to molting and exoskeleton change (chh, ctbs, gap65), stress and apoptosis (hsp40, hsp70, casp3), as well for detoxification (cyp6a18). In conclusion, this study shows that teflubenzuron can bioaccumulate in shrimps living in the littoral zone and at sublethal concentrations affects molecular mechanisms in non-hepatopancreatic tissue.

Development of anaesthetic protocols for lumpfish (Cyclopterus lumpus L.): Effect of anaesthetic concentrations, sea water temperature and body weight.

In recent years, use of lumpfish (Cyclopterus lumpus L.) as cleaner-fish to remove sea-lice have been chosen by many salmon farmers in Europe and Canada as an alternative to medical treatment, which has led to large scale production of lumpfish. At present, there is limited knowledge of how lumpfish respond upon anaesthesia, which anaesthetics and concentrations that are efficient and conditions for euthanasia. We have therefore tested and developed protocols for bath immersion for three commonly used anaesthetics metacaine (Finquel, buffered tricaine methanesulfonate, MS-222 and Tricaine Pharmaq), benzocaine (Benzoak vet) and isoeugenol (Aqui-S), determined concentration for normal and fast anaesthesia and evaluated safety margin for each condition. Also, a behavioral matrix has been developed. We have examined the effect of fish size (10-20 g, 200-400 g and 600-1300 g) and sea water temperature (6°C and 12°C). We found that 200 mg L-1 metacaine is an efficient dose for deep narcosis independently for fish size and temperature due to good safety margins with regards to both exposure times and doses. However, for many tasks lighter anaesthesia is sufficient, and then 100 mg L-1 metacaine can be used. Benzocaine is less efficient than metacaine, but can be used as anaesthetic of fish < 400 g. The optimal doses of benzocaine were 100-200 mg L-1 for small fish (10-20 g) and 200 mg L-1 for medium sized fish (200-400 g). For larger fish (> 600 g), benzocaine is not suitable. Isoeugenol cannot be recommended for full anesthesia of lumpfish. The conditions for lethal doses varied with chosen anaesthetic, fish size and temperature. For small fish (10-20 g), exposure to 1600 mgL-1 of metacaine in 10 minutes it lethal. Guided protocols for non-lethal anaesthesia will contribute to ensure safe treatment of lumpfish according to an ethical standard for good fish welfare.

Persistence and Stability of Teflubenzuron and Diflubenzuron When Associated to Organic Particles in Marine Sediment.

The persistence and stability of the oral administered anti salmon-lice drugs teflubenzuron and diflubenzuron were tested when associated to organic material as faecal particles from Atlantic salmon and medicated food pellets. This laboratory study was performed in seawater under aerobic conditions, at 7°C in the dark and showed that both compounds were remarkably persistent and stable since no significant reduction in the concentrations of flubenzurons in sediment were seen after 24 weeks. Therefore neither chemical or microbial degradation nor outwashing seems to be important pathways for these drugs to disappear from sediment under fish farms. Thus, it is more likely that the decrease of flubenzurons from marine sediments described in field investigations is caused by either bioturbation, resuspension of organic particles or a combination of these.

Transcriptional responses to teflubenzuron exposure in European lobster (Homarus gammarus).

Increasing use of pharmaceutical drugs to delouse farmed salmon raises environmental concerns. This study describes an experiment carried out to elucidate the molecular mechanisms of the antiparasitic drug teflubenzuron on a non-target species, the European lobster. Juvenile lobsters (10.3±0.9 mm carapace length) were fed two environmentally relevant doses of teflubenzuron, corresponding to 5 and 20% of a standard salmon medication (10 mg/kg day), termed low and high dose in this study. After 114 days of dietary exposure, whole-animal accumulation of teflubenzuron was determined. One claw from each animal was collected for transcriptional analysis. Overall, exposed animals showed low cumulative mortality. Six animals, two from the low dose treatment and four from the high dose, showed exoskeletal abnormalities (claw deformities or stiff walking legs). Residual levels of teflubenzuron in juvenile lobster were 2.7-fold higher in the high dose (282 ng/g) compared to the low dose treatment (103 ng/g). The transcriptional examination showed significant effects of teflubenzuron on 21 out of 39 studied genes. At the transcriptional level, environmentally relevant levels of the anti-salmon lice drug impacted genes linked to drug detoxification (cyp3a, cyp6a2, cyp302a, sult1b1, abcc4), cellular stress (hsp70, hsp90, chh), oxidative stress (cat, gpx3) and DNA damage (p53), as well as molting and exoskeleton regulation (chi3l1, ecr, jhl1, chs1, ctbs, gap65, jhel-ces1) in claw tissue (muscle and exoskeleton). In conclusion, teflubenzuron at sub-lethal levels can affect many molecular mechanisms in European lobster claws.

Distribution and persistence of the anti sea-lice drug teflubenzuron in wild fauna and sediments around a salmon farm, following a standard treatment.

The salmon louse (Lepeoptheirus salmonis) is a challenge in the farming of Atlantic salmon (Salmo salar). To treat an infestation, different insecticides are used like the orally administered chitin synthetase inhibitor teflubenzuron. The concentrations and distribution of teflubenzuron were measured in water, organic particles, marine sediment and biota caught in the vicinity of a fish farm following a standard medication. Low concentrations were found in water samples whereas the organic waste from the farm, collected by sediment traps had concentrations higher than the medicated feed. Most of the organic waste was distributed to the bottom close to the farm but organic particles containing teflubenzuron were collected 1100 m from the farm. The sediment under the farm consisted of 5 to 10% organic material and therefore the concentration of teflubenzuron was much lower than in the organic waste. Teflubenzuron was persistent in the sediment with a stipulated halflife of 170 days. Sediment consuming polychaetes had high but decreasing concentrations of teflubenzuron throughout the experimental period, reflecting the decrease of teflubenzuron in the sediment. During medication most wild fauna contained teflubenzuron residues and where polychaetes and saith had highest concentrations. Eight months later only polychaetes and some crustaceans contained drug residues. What dosages that induce mortality in various crustaceans following short or long-term exposure is not known but the results indicate that the concentrations in defined individuals of king crab, shrimp, squat lobster and Norway lobster were high enough shortly after medication to induce mortality if moulting was imminent. Considering food safety, saith and the brown meat of crustaceans contained at first sampling concentrations of teflubenzuron higher than the MRL-value set for Atlantic salmon. The concentrations were, however, moderate and the amount of saith fillet or brown meat of crustaceans to be consumed in order to exceed ADI is relatively large.

Mortality and deformities in European lobster (Homarus gammarus) juveniles exposed to the anti-parasitic drug teflubenzuron.

This study describes experiments carried out to examine effects of the antiparasitic drug teflubenzuron, used in delousing farmed salmon, on a non-target species, the European lobster (Homarus gammarus). Juvenile lobsters were fed two doses of teflubenzuron, 10 and 20mg/kg successively for 7 days corresponding to a standard medication of the fish (10mg/kg day) and twice the standard dose (20mg/kg day). Monitoring lasted 3 months to include at least one moulting period for all individuals. Cumulative mortality was higher in all replicates given medicated feed compared with the control group. Mean cumulative mortality for each dosing was 41 ± 13% for 10mg/kg and 38 ± 8% for 20mg/kg, i.e. no difference. Drug residue was analysed in all juveniles that died, in addition to 12 juveniles at day 8 and the first 12 surviving lobsters. A decline in concentration of teflubenzuron from over 8,000 ng/g (day 5) to 14 ng/g (day 70) was observed in the juveniles that died during the experiment. Twelve individuals that died contained 82 ng/g or less whereas the mean concentration in the first 12 lobsters that survived moulting was 152 ng/g. Following a single oral administration, the half-life of teflubenzuron in lobster was estimated to 3.4 days and the initial concentration (C0) to 515 ng/g at time t0. At the end of the study a considerable number of juvenile lobsters were observed with deformities in various organs; carapace, walking legs, cheliped, tail fan, abdomen and antenna. The occurrence of observed deformities varied from 0 to 15% in treated replicates and will most likely affect ability to locate and consume food (antenna, claw and walking legs), respiration (carapace) and ability to move/swim (walking legs, tail fan and abdomen). In total, the mortality and senescent damages were close to 50% in all replicates. Juveniles that survived medication without deformities however, moulted and increased in size at each moult equally well as the unmedicated controls.

Toxicological assessment of the anti-salmon lice drug diflubenzuron on Atlantic cod Gadus morhua.

Increasing use of the chitin synthesis inhibitor diflubenzuron against the ectoparasitic salmon louse Lepeophtheirus salmonis in marine aquaculture has raised concerns over its environmental impacts. This study evaluated how diflubenzuron affects Atlantic cod Gadus morhua, a fish species often found near Atlantic salmon Salmo salar farms, focusing on uptake kinetics and hepatic transcriptional responses. Two experiments were conducted, one time-series trial in which the fish were given a daily dose (3 mg kg-1 fish) of diflubenzuron for 14 d followed by a 3 wk depuration period, and one dose-response trial with increasing concentrations (3, 10 and 50 mg kg-1 fish). The highest diflubenzuron concentrations were found in the liver at Day 15. No detectable levels of diflubenzuron were found in liver or muscle 3 wk after the end of the treatment. At the molecular level, small effects of diflubenzuron treatment on gene transcription were observed. In the time-series experiment, the strongest effects were seen at Day 8, with 2 transcripts being upregulated (bclx2 and cpt1a) and 8 transcripts being downregulated (gstp1, gstm1, gstt1, ugt1a, nat2, cat, p53 and slc16a9a). Five transcripts (cyp3a, cpt1a, ptgs2, elovl5 and mapk1) responded significantly to diflubenzuron exposure in the dose-response experiment. This study shows that diflubenzuron can be taken up by Atlantic cod, that it is rapidly cleared from the body and that when present this pharmaceutical causes only small effects on the expression of genes involved in detoxification pathways. Taken together, our data suggest that accumulated diflubenzuron at the levels studied would have a relatively small effect on wild Atlantic cod.

Antimicrobial susceptibility of Francisella noatunensis subsp. noatunensis strains isolated from Atlantic cod Gadus morhua in Norway.

A total of 30 isolates of Francisella noatunensis subsp. noatunensis isolated from Atlantic cod Gadus morhua L. were tested for susceptibility, in the form of minimal inhibitory concentration (MIC) values, against the following antibacterial agents: flumequine, oxolinic acid, ciprofloxacin, florfenicol, oxytetracycline, erythromycin, streptomycin sulphate, trimetoprim/sulphadiazine and rifampin. All the isolates had a low susceptibility to oxytetracycline, trimetoprim/sulphadiazine (Tribrissen®), erythromycin, ciprofloxacin and streptomycin with MIC values of 64, 64 to 128, 16, 8 and 32 to 128 µg ml-1, respectively. The strains were, on the other hand, susceptible to florfenicol, oxolinic acid, flumequine and rifampin with MIC values of 0.5, 0.25, 0.25 and 0.25 to 1 µg ml-1, respectively.