The effects of emamectin benzoate or ivermectin spiked sediment on juvenile American lobsters (Homarus americanus).

This study examined the effects of a range of ½-log concentrations of emamectin benzoate (commercially applied as SLICE®) and ivermectin (commercially applied as IVOMEC®) on juvenile American lobster, Homarus americanus. Phase I of the research assessed acute (up to 4 days) and chronic (30-day) toxicity of sediment dosed with the active ingredients emamectin benzoate (EMB) formulated as SLICE® or ivermectin (IVM) formulated as IVOMEC® at various nominal concentrations (EMB: 15, 48, 150, 475 and 1500 ng g-1 wet sediment; IVM: 3, 9.5, 30, 95 and 300 ng g-1 wet sediment) on juvenile Atlantic lobster (stages IV). Phase II evaluated sublethal effects (e.g., growth, moulting success) of all lobster surviving past the 30 day exposure period, over an additional 41 days. Chemical analysis of EMB and IVM in sediment samples from the exposure tanks revealed a strong linear association (R2 values 0.99 and 0.98 for EMB and IVM, respectively) between nominal dose and measured concentration of compound. EMB exposure concentrations at very high levels (≥ 343.3 ng g-1) were acutely toxic to juvenile lobster such that 100% of lobsters had died after 13 days of exposure. The maximum cumulative mortality of lobsters exposed to the highest concentrations of EMB and IVM was 100% after 10 days and 25 days, respectively. The 10-day LC50 estimates (±â€¯95% CI) for EMB and IVM were 250.23 ±â€¯90.4 and 212.14 ±â€¯202.64 ng g-1, respectively. Using abnormal behaviour as an indicator, the 15-day EC50 estimates (±â€¯95% CI) for EMB and IVM were 96.19 ±â€¯51.42 and 15.82 ±â€¯6.93 ng g-1, respectively. The NOEC (no observed effect concentration) for abnormal behaviour was 0.0 ng g-1 for each product and the LOEC (lowest observed effect concentration) was 8.8 and > 3.0 ng g-1 for EMB and IVM, respectively. Observations on sublethal effects included delayed moulting to stage VI and reduced growth at higher exposure concentrations for both therapeutants. Using failure to moult to stage V or VI as an indicator, the 15-day EC50 estimates (±â€¯95% CI) for EMB and IVM were 32.72 ±â€¯18.26 and 14.00 ±â€¯12.43 ng g-1, respectively. The NOEC for failure to moult to stage V only was 343.3 and 14.7 ng g-1 for EMB and IVM, respectively. Whereas, the LOEC was 1066.7 and > 61.0 ng g-1 for EMB and IVM, respectively. The concentrations of EMB and IVM tested in the present study were acutely toxic to juvenile lobster exposed to the highest dosages (343.3 and 1066.7 ng EMB g-1 and 61.0 and 300.0 ng IVM g-1). There was significant evidence of chronic toxicity, longer exposure increased mortality with LT50 values decreasing with increasing test material concentration.

Distinctive metabolite profiles in in-migrating Sockeye salmon suggest sex-linked endocrine perturbation.

The health of Skeena River Sockeye salmon (Onchorhychus nerka) has been of increasing concern due to declining stock returns over the past decade. In the present work, in-migrating Sockeye from the 2008 run were evaluated using a mass spectrometry-based, targeted metabolomics platform. Our objectives were to (a) investigate natural changes in a subset of the hepatic metabolome arising from migration-associated changes in osmoregulation, locomotion, and gametogenesis, and (b) compare the resultant profiles with animals displaying altered hepatic vitellogenin A (vtg) expression at the spawning grounds, which was previously hypothesized as a marker of xenobiotic exposure. Of 203 metabolites monitored, 95 were consistently observed in Sockeye salmon livers and over half of these changed significantly during in-migration. Among the most dramatic changes in both sexes were a decrease in concentrations of taurine (a major organic osmolyte), carnitine (involved in fatty acid transport), and two major polyunsaturated fatty acids (eicosapentaenoic acid and docosahexaenoic acid). In females, an increase in amino acids was attributed to protein catabolism associated with vitellogenesis. Animals with atypical vtg mRNA expression demonstrated unusual hepatic amino acid, fatty acid, taurine, and carnitine profiles. The cause of these molecular perturbations remains unclear, but may include xenobiotic exposure, natural senescence, and/or interindividual variability. These data provide a benchmark for further investigation into the long-term health of migrating Skeena Sockeye.

Development and validation of protocols to differentiate PCB patterns between farmed and wild salmon.

Polychlorinated biphenyl (PCB) congener patterns based on full congener PCB analyses of three farmed and five wild species of salmon from coastal British Columbia, Canada are compared using principal components analysis (PCA) and the best fit linear decomposition of the observed PCB composition in terms of Aroclor 1242, 1254, and 1260 end-members. The two complementary analysis methods are used to investigate congener composition pattern differences between species, trophic levels, feeding preferences, and farmed or wild feeding regimes, with the intent of better understanding PCB processes in both salmon and salmon consumers. PCA supports classification of PCB congeners into nine groups based on a) structure activity groups (SAG) related to the bioaccumulation potential in fish-eating mammals, b) Cl number, and c) the numbers of vicinal meta- and para-H. All three factors are needed to interpret congener distributions since SAGs by themselves do not fully explain PCB distributions. Farmed salmon exhibit very similar congener patterns that overlap the PCA and Aroclor composition of their food, while wild salmon separate into two distinct groups, with chinook and "coastal" coho having higher proportions of the higher chlorinated, Aroclor 1260 type, nonmetabolizable congeners, and chum, pink, sockeye, and "remote" coho having higher proportions of the lower chlorinated, more volatile and metabolizable Aroclor 1242 type, congeners. Wild chinook have the highest PCB and toxic equivalent (TEQ) concentrations, and the highest proportions of A1254 A1260, and PCB congeners in the most refractory SAG. Because both "coastal" and "remote" coho groups are likely to be consuming prey of similar size and trophic level, the PCB delivery mechanism (e.g., atmosphere vs runoff) apparently has more influence on the salmon PCB profile than biotransformation, suggesting that the wild chinook PCB profile is determined by feeding preference. Overall, wild salmon distributions primarily relate to trophic level, feeding preferences, and longevity, while metabolism appears at most a minor factor. The new classification protocol takes better advantage of individual congener PCB analyses and provides a better framework for understanding the PCB distributions in salmon and, potentially, the movement of individual PCB congeners through marine food chains than previous classification schemes.

Mercury and other trace elements in farmed and wild salmon from British Columbia, Canada.

The present study reports measured levels of Hg and other trace elements in commercial salmon feed; farmed Atlantic, coho, and chinook salmon (n = 110); and wild coho, chinook, chum, sockeye, and pink salmon (n = 91). Metal concentrations in farmed and wild salmon from British Columbia, Canada, were relatively low and below human health consumption guidelines. Methylmercury in all salmon samples (range, 0.03-0.1 microg/g wet wt) were below the 0.5 microg/g guideline set by Health Canada. Negligible differences in metal concentrations were observed between the various species of farmed and wild salmon. Metal concentrations generally were higher in commercial salmon feed compared to farmed salmon. Mercury showed slight bioaccumulation potential in farmed salmon, with biomagnification factors (BMFs) ranging between 0.8 and 1.9. Other metals, such as Cd, Pb, and Ni, exhibited biodilution, with BMFs of much less than one. The relatively low degree of biomagnification of metals observed in farmed salmon likely resulted from the combination of low gastrointestinal absorption efficiency, negligible transfer to muscle tissue relative to other compartments, and a high degree of growth dilution in these fish. Human dietary exposure calculations indicate intakes of Hg, Cd, Pb, Cu, As, and Ni via farmed and wild British Columbia salmon are a relatively small percentage of total intakes (0.05-32%) compared to other Canadian foodstuffs, such as fruits, vegetables, chicken, and beef (68-99%). Although total dietary exposure of Cd, Pb, and Cu approached provisional tolerable daily intake levels, the contribution from British Columbia salmon was less than 2%. Our findings indicate farmed and wild British Columbia salmon remain a safe source of omega-3 highly unsaturated fatty acid intake for cardioprotective and, possibly, other health benefits.

Evidence of disruption in estrogen-associated signaling in the liver transcriptome of in-migrating sockeye salmon of British Columbia, Canada.

The health of sockeye (Oncorhynchus nerka) salmon stocks is of increasing concern; reflecting both a sentinel of human-impacted aquatic environments and as a key fishery for British Columbia, Canada. The spawning migration of Pacific sockeye salmon represents a critical life stage where significant demands are made on animal biology and important BC fisheries are linked to this migration in the Skeena and Fraser River watersheds. These watersheds present very different environments; the former being sparsely populated with little industrial impact, while the latter flows through highly-populated areas. The present study used quantitative real-time PCR analysis of adult sockeye salmon from four 2008 stocks [Fulton River and Pinkut Creek (Skeena) and Weaver Creek and Harrison River (Fraser)] to evaluate ten hepatic gene transcripts associated with reproduction, stress, energy metabolism, and exposure to environmental contaminants. Dynamic changes in mRNA abundance were observed in Fulton River stock animals from the Skeena River mouth to the spawning ground which reflect the physiological demands of in-river migration and reproductive maturation. Inter-stock comparisons of migrants at spawning grounds demonstrated a marked difference in the sex-specific gene hepatic gene expression profiles. Our original hypothesis was that a greater diversity in mRNA profiles is associated with watersheds with higher human impact. However, our observations contradict this posit. Skeena males and females displayed poor definition in their molecular profiles between sexes while the Fraser River fish had very distinctive sex differences that were consistent with the previous year's migration. The genetic sex distribution and ratio of milt versus roe production did not differ between the Skeena and Fraser River spawning site fish. However, a significant percentage of Skeena animals displayed marked discordance of these characteristics with gender-specific hepatic mRNA profiles implying that an alteration in estrogen-mediated signaling has occurred. Continued geospatial and longitudinal assessments will help determine to what extent the dynamic molecular biology of late life-stage sockeye salmon reflects natural variation or modulation by anthropogenic causative agents.

Biological effects of the anti-parasitic chemotherapeutant emamectin benzoate on a non-target crustacean, the spot prawn (Pandalus platyceros Brandt, 1851) under laboratory conditions.

The potential impact of commercial salmon aquaculture along the coast of British Columbia on the health of non-target marine wildlife is of growing concern. In the current initiative, the biological effects on gene expression within spot prawn (Pandalus platyceros) exposed to the sea lice controlling agent, emamectin benzoate (EB; 0.1-4.8 mg/kg sediment), were investigated. A mean sediment/water partitioning coefficient (K(p)) was determined to be 21.81 and significant levels of EB were detected in the tail muscle tissue in all exposed animals. Animals selected for the experiment did not have eggs and were of similar weight. Significant mortality was observed within 8 days of EB treatment at concentrations between 0.1 and 0.8 mg/kg and there was no effect of EB on molting. Twelve spot prawn cDNA sequences were isolated from the tail muscle either by directed cloning or subtractive hybridization of control versus EB exposed tissues. Three of the transcripts most affected by EB exposure matched sequences encoding the 60S ribosomal protein L22, spliceosome RNA helicase WM6/UAP56, and the intracellular signal mediator histidine triad nucleotide binding protein 1 suggesting that translation, transcription regulation, and apoptosis pathways were impacted. The mRNA encoding the molting enzyme, ß-N-acetylglucosaminidase, was not affected by EB treatment. However, the expression of this transcript was extremely variable making it unsuitable for effects assessment. The results suggest that short-term exposure to EB can impact biological processes within this non-target crustacean.

Flesh residue concentrations of organochlorine pesticides in farmed and wild salmon from British Columbia, Canada.

The present study reports measured levels of organochlorine pesticides (OCPs) in commercial salmon feed (n = 8) and farmed Atlantic, coho, and chinook salmon (n = 110), as well as wild coho, chinook, chum, sockeye, and pink salmon (n = 91). Flesh residue concentrations (ng/g wet weight) of dichlorodiphenyltrichloroethanes (DDTs), hexachlorocyclohexanes (HCHs), chlordanes, chlorobenzenes (CBz) and cyclodiene pesticides (e.g., dieldrin, mirex) were 2 to 11 times higher (p < 0.05) in farmed salmon compared with wild salmon. Concentrations were positively correlated with flesh lipid levels. Farmed Atlantic salmon (12-15% lipid) typically exhibited the greatest OCP burdens compared with other salmon species. However, when expressed on a lipid weight basis, concentrations of OCPs (ng/g lipid weight) in wild salmon, in many cases, exceeded those levels in farmed salmon. Observed interspecies and site-specific variations of OCP concentrations in farmed and wild salmon may be attributed to divergent life history, prey/feed characteristics and composition, bioenergetics, or ambient environmental concentrations. Calculated biomagnification factors (BMF = C(F)/C(D), lipid wt) of OCPs in farmed salmon typically ranged between two and five. Biomagnification of chemicals such as DDTs, chlordanes, and mirex was anticipated, because those compounds tend to exhibit high dietary uptake and slow depuration rates in fish because of relatively high octanol-water partition coefficients (K(OW)s > 105). Surprisingly, less hydrophobic pesticides such as hexachlorocyclohexanes and endosulfans (K(OW) s < 105) consistently exhibited a high degree of biomagnification in farmed salmon species (BMFs > 5). This is contrary to previous laboratory and field observations demonstrating fish BMFs less than 1 for low K(OW) chemicals, because of efficient respiratory elimination of those compounds via gills. The results suggest that ambient seawater concentrations and bioconcentration-driven accumulation may play a key role in the bioaccumulation of these relatively more water-soluble contaminants in farmed salmon. Finally, OCP exposure through consumption of British Columbian salmon is found to be low relative to United States national average per capita total exposure levels and provisional tolerable daily intakes.

Tissue residue concentrations of organohalogens and trace elements in adult Pacific salmon returning to the Fraser River, British Columbia, Canada.

We report measured concentrations of organohalogens and trace elements in muscle and eggs of returning wild Pacific sockeye and chinook salmon during their 2007 migration through the Fraser River watershed in Canada. Chemical analyses revealed the presence of ppb to ppm levels of a wide variety of contaminants in these fish, including polychlorinated biphenyls (PCBs); polychlorinated dibenzo-p-dioxins (PCDDs); polychlorinated dibenzofurans (PCDFs); polybrominated diphenyl ethers (PBDEs); organochlorine pesticides (OCPs) such as DDTs, hexachlorocyclohexanes (HCHs), octachlorostyrene, and cyclodienes; and Hg, As, Cd, Pb, and several other trace elements. Body weights and flesh lipid contents declined during upstream migration, resulting in significantly higher (p < 0.05) lipid-normalized concentrations of lipophilic organohalogens (PCBs, PCDD/Fs, pesticides) in those spawning salmon. Postmigration magnification factors (MFs) of organohalogens (0.1-10) were comparable to previous observations and model predictions. MFs generally increased with increasing hydrophobicity (K(OW)). For example, MFs of tetra- and pentachlorobenzenes and HCH isomers (log K(OW) range: 3.8-5) were relatively low (between 0.1 and 1.7) compared with those of more lipophilic compounds (log K(OW) > 6) such as PCBs, DDTs, and mirex (MFs between 5 and 10). Lipid-normalized muscle:egg ratios in female salmon, which varied between 0.1 and 8, also exhibited a positive relationship with chemical K(OW). The results indicate that lipophilic compounds (K(OW) > 10(6)) can be magnified in flesh lipids of Pacific salmon during spawning migration, but maternal transfer kinetics (deposition to eggs) of those chemicals are relatively slow compared with less hydrophobic compounds. 2,3,7,8-TCDD toxic equivalents (ΣTEQs) in eggs of these spawning salmon, calculated using WHO toxic equivalency factors (WHO-TEFs) for fish health, in some cases exceeded the 0.3 pg·g(-1) threshold level associated with 30% salmonid egg mortality, indicating the potential for reproductive impacts in Fraser River salmon populations.

DDT in endangered Galapagos sea lions (Zalophus wollebaeki).

We characterize for the first time the presence of DDT and its metabolites in tropical Galapagos sea lions (Zalophus wolleabeki). ∑DDT concentrations in Galapagos sea lion pups sampled in 2005 and 2008 ranged from 16 to 3070 µg/kg lipid. Concentrations of ∑DDT in pups in 2008 averaged 525 µg/kg lipid and were 1.9 times higher than that (281 µg/kg lipid) detected in pups in 2005. These concentrations are lower than those reported in many pinnipeds elsewhere, comparable to those in Hawaiian monk seals, and higher than those in southern elephant seals. The health risk characterization showed that 1% of the male pups exceeded the p,p'-DDE toxic effect concentration associated with anti-androgenic effects reported in rats. The findings provide preliminary guidance on the relationship between DDT use and ecological impacts, serving as a reference point against which possible future impact of tropical DDT use can be assessed.

Gene expression profiling and environmental contaminant assessment of migrating Pacific salmon in the Fraser River watershed of British Columbia.

The health and physiological condition of anadromous salmon is of concern as their upriver migration requires navigation of human-impacted waterways and metabolism of stored energy reserves containing anthropogenic contaminants. Such factors may affect reproductive success of fish stocks. This study investigates chemical contaminant burdens and select gene expression profiles in Pacific Sockeye (Oncorhynchus nerka) and Chinook (Oncorhynchus tshawytscha) salmon which traverse the Fraser River watershed during their spawning migration. Chemical analyses of muscle tissue and eggs of salmon collected from the lower Fraser River (pre-migration) and from upstream spawning grounds (post-migration) during the 2007 migration revealed the presence of numerous chemical contaminants, including PCBs, dioxins/furans, pesticides, and heavy metals. However, muscle tissue residue concentrations were well below human health consumption guidelines and 2,3,7,8 TCDD toxic equivalents (SigmaTEQs) in salmon eggs, calculated using WHO toxic equivalency factors (WHO-TEFs) for fish health, did not exceed the 0.3pgg(-1) wet weight toxicological threshold level previously associated with 30% egg mortality in salmon populations. Quantitative real-time PCR probes were generated and used to assess differences in abundance of key mRNA transcripts encoding nine gene products associated with reproduction, stress, metal toxicity, and exposure to environmental contaminants. Gene expression profiles were characterized in liver and muscle tissue of pre- and post-migration Sockeye and Chinook salmon. The results of stock-matched animals indicate that dynamic changes in mRNA levels occur for a number of genes in both species during migration and suggest that Sockeye salmon exhibit a greater level of biological stress compared to the Chinook salmon population. Using a male-specific genotypic marker, we found that out of the 154 animals examined, one Sockeye was genotypically male but phenotypically female. This individual's gene expression profile in liver and muscle was reminiscent of, but not identical to, the female expression profile. These studies provide the first glimpse of the dynamic yet common nature of changes in the transcriptome that are shared between species during in-migration and highlight differences that may relate to population success. Continued longitudinal assessment will further define the association between contaminant burden, physiological stress, and modulation of gene expression in migrating Pacific salmon.

Use of terrestrial based lipids in aquaculture feeds and the effects on flesh organohalogen and fatty acid concentrations in farmed Atlantic salmon.

Consumption of salmon, wild or farmed, has been encouraged by many scientists and by national and international health organizations due to the potential health benefits associated with their high contents of omega-3 (n-3) highly unsaturated fatty acids (n-3 HUFAs). In 2004, there was increased public concern regarding the safety of farmed Atlantic salmon following the publication of several studies that indicated higher levels of organohalogens in their flesh relative to those noted in the flesh of wild Pacific salmon. Farmed salmon obtain most of these contaminants from the consumption of marine fish oil (MFO) present in salmon feed. In both a laboratory feeding trial and an on-farm field study, partial replacement of MFO in aquaculture feeds with economical and abundant lipids of terrestrial origin resulted in farmed Atlantic salmon with reduced flesh polychlorinated biphenyl and polychlorinated dibenzodioxin and furan concentrations. Flesh levels of n-3 HUFAs (g/(100 g serving)) were lower in farmed Atlantic salmon fed diets with alternative lipids relative to farmed salmon fed more traditional MFO-based diets. However, the former salmon were found to have higher flesh levels of n-3 HUFAs and also similar or lower flesh levels of organic contaminants than some species of market-size wild Pacific salmon. These findings showthat consumption of either farmed Atlantic salmon or wild Pacific salmon can meet recommended weekly n-3 HUFA levels with minimal concurrent intake of flesh organohalogens.