Exposure of female lobsters to sediments spiked with emamectin benzoate: Effects on eggs and larvae early development.

The salmon aquaculture industry is an important economic activity established on both the west and east coast of Canada. To control sea lice infestations, in-feed products like emamectin benzoate (EMB) are widely used. Due to its low solubility and persistence EMB can accumulate in marine sediments and be potentially bioavailable to non-target organisms from months to years. The American lobster (Homarus americanus) is a key species in the Northwest Atlantic with high economic and ecological value. It may be exposed to therapeutants considering lobster habitats overlap with aquaculture locations requiring a better understanding of the potential impact of these therapeutants through varied pathways of exposure. In this study, we investigated the exposure of gravid female lobsters to EMB spiked sediment to mimic the likely presence of these females at aquaculture sites for a 10-day period. We completed testing by assessing EMB effects on adult molting and quality, embryo hatching rates, and larval offspring quality and larval molting. Our results show that a single, 10-day exposure of ovigerous females to EMB concentrations higher than environmentally relevant values did not affect females or their offspring.

Considerations and data update for the inference of environmental quality standards for two avermectins widely used in salmon aquaculture.

We have updated and reviewed toxicity data for Emamectin benzoate (EMB) and Ivermectin (IVER), two in-feed drugs used to treat sea lice in farmed Atlantic salmon, and inferred new Environmental Quality Standards (EQS) using a deterministic approach or Species Sensitivity Distributions (SSDs) based on available data. We used a SSD model averaging approach and inferred a water acute EQS value of 24.9 ng/L (SSD) for EMB, while previously established chronic water EQS of 0.17 ng/L and sediment benthic EQS of 131 ng/kg dry weight remained unchanged. For IVER, both a water acute EQS of 8.04 ng/L and a chronic water EQS of 3.98 ng/L were inferred using SSDs as well as a benthic EQS of 290 ng/kg dry weight using a deterministic approach. In light of the lack of solubility and tendency of both avermectins to sorb to material benthic EQSs remain the most relevant value to consider for regulators.

The persistence of emamectin benzoate in marine sediments with different organic matter regimes, temperature conditions, and antibiotic presence.

The objectives of this 318-day study are to determine half-lives of the anti-sea lice medication emamectin benzoate (EMB) under conditions present in sediments at aquaculture sites and document the degradation of EMB into its main metabolite desmethyl emamectin benzoate (DES). Tested conditions include different matrix types (sand, mud), two temperatures (4, 10 degrees), organic matter presence (fish feed waste and feces), and the presence of oxytetracycline. We document a transformation ratio of EMB to DES of 0.16 to 4.4 % and show that the co-presence of oxytetracycline increases EMB calculated half-lives to values >6000 days for mud matrices. EMB incubated in high organic enrichment regimes was not observed to degrade at 4 degrees. Multivariate analyses show interactions between sediment conditions (matrix, temperature, organic matter [OM], oxytetracycline) influence EMB persistence and DES:EMB ratios. Ranges of EMB half-lives and information on metabolites can be used to anticipate potential effects on marine communities.

Relationship between in feed drugs, antibiotics and organic enrichment in marine sediments at Canadian Atlantic salmon aquaculture sites.

The presence of in-feed anti-sea lice drugs and their relationship with organic enrichment is poorly understood in sediment surrounding salmon farms. Using data from an aquaculture monitoring program (2018-2020), we describe this relationship at ten sites in four Canadian provinces. Three anti-sea lice pesticides (lufenuron, teflubenzuron, emamectin benzoate and metabolite desmethyl emamectin benzoate), and one antibiotic (oxytetracycline) were detected. Concentrations were often below limits of quantification. Values are also lower than those reported in other aquaculture salmon-producing countries. Highest concentrations, along with organic enrichment, were observed ~200 m of cages with lower concentrations detected up to 1.5 km away. Most samples had at least two drugs present: 75.2 % (British Columbia), 91.4 % (Newfoundland), and 54.8 % (New Brunswick/Nova Scotia) highlighting the potential for cumulative effects. Emamectin benzoate and oxytetracycline were detected four and three years respectively after last known treatments, demonstrating the need for research on overall persistence of compounds.

Metabolization of emamectin benzoate into desmethyl emamectin benzoate in spiked marine sediments.

Emamectin benzoate (EMB) (4″deoxy- 4″-epi-methylaminoavermectin) is a pesticide developed to control pests on various crops, and in forestry. It is also used in salmon aquaculture to control sea lice infestations as an in-feed therapeutant. Little is known about EMB metabolites and potential associated toxicities in marine sediments. In this study, we used natural marine sediments (sand and mud) fortified at an EMB concentration of 60 ppb (wet weight). Results show an almost immediate transformation of EMB to Desmethyl EMB (DES) with no increased rates of metabolization when stored sediment samples were incubated for up to 16 h. The transformation ratio of EMB to DES represented between 0.16 and 0.39% of EMB; values are lower than what has been observed in fish tissue. Data suggest that DES is generated through both abiotic (tested after autoclaving marine sediments) and biological processes. Further work on freshly sampled marine sediments with EMB deposits, different organic regimes, and a detailed assessment of active bacterial communities are necessary to better evaluate the EMB to DES rate of transformation around aquaculture sites.

Species sensitivity distributions for two widely used anti-sea lice chemotherapeutants in the salmon aquaculture industry.

The main objective of the present study is to construct acute aquatic species sensitivity distributions (SSD) and generate proposed HC5 values (i.e. the hazardous concentration for which 5 % of species are affected or potentially affected) for two aquaculture anti-sea lice bath pesticides, azamethiphos, and hydrogen peroxide. These values could be used as the basis for the establishment of environmental quality standards (EQS). We have generated SSDs and inferred HC5 values for mortality and sublethal endpoints using LC50, EC50, and NOEC/LOEC data points separately and for each bath pesticide. Through the examination of literature data on the toxicity of both compounds, we opted to use tests with limited exposure times to ensure environmental relevance for bath pesticides. We also separated life stages for some of the sensitive taxa to account for differences in sensitivities and risk of exposure. The resulting threshold concentrations in environmental seawater are 0.10 µg/L for azamethiphos and 0.15 mg/L for hydrogen peroxide. These suggested azamethiphos and hydrogen peroxide thresholds are comparable to some previously reported EQS values. Further considerations need to be included in how to better use these thresholds in a regulatory context in relation to dispersion patterns. It is also clear that delayed mortality and sublethal effects documented in the literature require further study to fully anticipate the environmental risks of using these two bath pesticides.

Assessing trace-elements as indicators of marine finfish aquaculture across three distinct Canadian coastal regions.

Several trace-elements have been identified as indicators of finfish aquaculture organic enrichment. In this study, sediment sampling at finfish farms was completed as part of an Aquaculture Monitoring Program in three distinct Canadian regions. Despite diverse datasets, multivariate analyses show a consistent clustering of known direct (Cu and Zn) and indirect (Cd, Mo and U) tracers of aquaculture activities with sediment organic matter (OM) and/or total dissolved sulfides concentrations. OM content was also a predictor of Cu, Zn, Mo and U concentrations according to decision tree analyses. Distance from cages did not emerge as a strong driver of differences among sampling points; however, a tendency towards negative associations is clear especially for Zn. Enriched stations as determined after geochemical normalization were mostly localized within 150 m of net-pens. Selected trace-elements (in particular Zn) can be useful indicators of aquaculture organic enrichment in different ecosystems and valuable tools for monitoring programs.

Early life traits of farm and wild Atlantic salmon Salmo salar and first generation hybrids in the south coast of Newfoundland.

This study examined fertilization rates, survival and early life-trait differences of pure farm, wild and first generation (F1) hybrid origin embryos after crossing farm and wild Atlantic salmon Salmo salar. Results show that despite a trend towards higher in vitro fertilization success for wild females, differences in fertilization success in river water are not significantly different among crosses. In a hatchery environment, wild females' progeny (pure wild and hybrids with wild maternal parent) hatched 7-11 days earlier than pure farm crosses and hybrids with farm maternal parents. In addition, pure wild progeny had higher total lengths (LT ) at hatch than pure farm crosses and hybrids. Directions in trait differences need to be tested in a river environment, but results clearly show the maternal influence on early stages beyond egg-size differences. Differences in LT were no longer significant at 70 days post hatch (shortly after the onset of exogenous feeding) showing the need to investigate later developmental stages to better assess somatic growth disparities due to genetic differences. Higher mortality rates of the most likely hybrids (farm female × wild male hybrids) at egg and fry stages and their delayed hatch suggest that these F1 hybrids might be less likely to survive the early larval stages than wild stocks.

Sperm traits in farmed and wild Atlantic salmon Salmo salar.

Differences in sperm metabolism and morphology between wild and non-local farmed Atlantic salmon Salmo salar were assessed by measuring metabolic enzyme activities and length of sperm flagella. No differences were observed between wild and farmed S. salar sperm with regards to cell counts or any of the biochemical variables assessed. Flagella of sperm cells were significantly longer in wild than farmed S. salar; however, this did not result in higher energy levels or different fertilization rates.

Effects of tebufenozide on some aspects of lake trout (Salvelinus namaycush) immune response.

Tebufenozide is a nonsteroid ecdysone agonist that causes premature and incomplete molting in Lepidoptera and is used on crops and in forest spray programs. Lake trout (Salvelinus namaycush) were exposed to one pulse of 0.25 ppm tebufenozide every 3 days (12 pulses in total). Cell ratios and respiratory burst responses of circulating white blood cells (WBCs) as well as head-kidney (HK) WBCs were investigated by using cell observation, Nitro-Blue Tetrazolium tests, and flow cytometry. Endpoints studied suggest a difference in sensitivity between HK and circulating blood cells to tebufenozide and show a stimulation of fish cell function and changes in percentages of cell types. Responses are not associated with a strong stress response as highlighted by the absence of effect on cortisol and blood protein levels. These results and tebufenozide persistence in water warrant further studies on pesticide impact on fish immunity when used on crops or in forest spray programs near lakes and streams.

Are metal mining effluent regulations adequate: identification of a novel bleached fish syndrome in association with iron-ore mining effluents in Labrador, Newfoundland.

Water quality guidelines for industrial effluents are in place in many countries but they have generally evolved within a limited ecotoxicological framework. Effluents from iron-ore mines have traditionally been viewed by regulatory bodies as posing little or no risk to the aquatic environment. However, it was recently reported that lake trout taken from a large iron-ore contaminated Lake in Labrador (Wabush Lake) had elevated levels of DNA oxidative damage and were markedly depleted in levels of vitamin A (Payne et al., 1998) in comparison with fish from a Lake (Shabogamo Lake) receiving lesser levels of effluents. Through further observations, it has now been established that the lake trout in Wabush Lake are commonly affected with a marked skin bleaching syndrome in comparison with fish in Shabogamo Lake and a nearby Lake (Ashuanipi) which does not receive effluents. To the authors' knowledge such a syndrome which is characterized by marked reduction in skin pigmentation and overall increase in skin whitening has not been reported before in any fish population in association with contamination. Preliminary information for liver histopathological and blood cell differences have also been obtained in fish in Wabush Lake in comparison with Ashuanipi Lake. It has also been observed through studies on phosphatidyl liposomes that iron-ore leachate contains redox-active material (iron but possibly other transition metals) that has considerable potential for causing oxidative damage to cellular constituents. Using the weight of evidence approach it is indicated that iron-ore effluents may pose more of a risk to the aquatic environment than traditionally considered by regulatory agencies.

Iron ore mines leachate potential for oxyradical production.

The ecotoxicological effects of mining effluents is coming under much greater scrutiny. It appears necessary to explore possible health effects in association with iron ore mining effluents. The present results clearly demonstrate that iron-ore leachate is not an inert media but has the potential to induce lipid peroxidation. Peroxidation was assessed by measuring oxygen consumption in the presence of a reducing agent such as ascorbate or NADPH and a chelator such as EDTA. Labrador iron ore is an insoluble complex crystalline material containing a mixture of metals (Fe, Al, Ti, Mn, Mg,ellipsis, ) in contrast to the iron sources used for normal lipid peroxidation studies. The metal of highest percentage is iron (59. 58%), a metal known to induce oxyradical production. Iron ore powder initiated ascorbic acid-dependent lipid peroxidation (nonenzymatic) in liposomes, lipids extracted from rat and salmon liver microsomes, and intact salmon liver microsomes. It also revealed an inhibitory effect of NADPH-dependent microsomes lipid peroxidation as well as on NADPH cytochrome c reductase activity. However, nonenzymatic peroxidation in rat liver microsomes was not significantly inhibited. Cytochrome P450 IA1- and IIB1-dependent enzymatic activities as well as P450 levels were not affected. The inhibition could be due to one of the other components of iron ore leachate (Mn, Al,ellipsis, ). These effects of iron-ore leachate indicate that a potential toxicity could be associated with its release into lakes. Further studies are necessary to explore in vivo effects on aquatic animals.