DNA damage and health effects in juvenile haddock (Melanogrammus aeglefinus) exposed to PAHs associated with oil-polluted sediment or produced water.

The research objective was to study the presence of DNA damages in haddock exposed to petrogenic or pyrogenic polyaromatic hydrocarbons (PAHs) from different sources: 1) extracts of oil produced water (PW), dominated by 2-ring PAHs; 2) distillation fractions of crude oil (representing oil-based drilling mud), dominated by 3-ring PAHs; 3) heavy pyrogenic PAHs, mixture of 4/5/6-ring PAHs. The biological effect of the different PAH sources was studied by feeding juvenile haddock with low doses of PAHs (0.3-0.7 mg PAH/kg fish/day) for two months, followed by a two-months recovery. In addition to the oral exposure, a group of fish was exposed to 12 single compounds of PAHs (4/5/6-ring) via intraperitoneal injection. The main endpoint was the analysis of hepatic and intestinal DNA adducts. In addition, PAH burden in liver, bile metabolites, gene and protein expression of CYP1A, GST activity, lipid peroxidation, skeletal deformities and histopathology of livers were evaluated. Juvenile haddock responded quickly to both intraperitoneal injection and oral exposure of 4/5/6-ring PAHs. High levels of DNA adducts were detected in livers three days after the dose of the single compound exposure. Fish had also high levels of DNA adducts in liver after being fed with extracts dominated by 2-ring PAHs (a PW exposure scenario) and 3-ring PAHs (simulating an oil exposure scenario). Elevated levels of DNA adducts were observed in the liver of all exposed groups after the 2 months of recovery. High levels of DNA adduct were found also in the intestines of individuals exposed to oil or heavy PAHs, but not in the PW or control groups. This suggests that the intestinal barrier is very important for detoxification of orally exposures of PAHs.

Hepatobiliary Analyses Suggest Chronic PAH Exposure in Hakes (Urophycis spp.) Following the Deepwater Horizon Oil Spill.

Prior to the Deepwater Horizon oil spill, we lacked a comprehensive baseline of oil contamination in the Gulf of Mexico's sediments, water column, and biota. Gaps in prespill knowledge limit our ability to determine the aftereffects of the Deepwater Horizon blowout or prepare to mitigate similar impacts during future oil spill disasters. We examined spatiotemporal differences in exposure to and metabolism of polycyclic aromatic hydrocarbons (PAHs) in 2 hake species (Urophycis spp.) to establish a current baseline for these ecologically important, abundant, and at-risk demersal fishes. Gulf hake (Urophycis cirrata) and southern hake (Urophycis floridana) were collected throughout the Gulf of Mexico during extensive longline surveys from 2012 to 2015. Analyses of biliary PAH metabolites and liver PAH concentrations provided evidence of exposures to di- and tricyclic compounds, with the highest concentrations measured in the northern Gulf of Mexico. Species-specific differences were not detected, but temporal trends observed in biliary PAHs suggest a decrease in acute exposures, whereas increasing liver PAHs suggest chronic exposures marked by greater assimilation than metabolism rates. To our knowledge, the present study provides the first multitissue contaminant analyses, as well as the most exhaustive biometric analyses, for both gulf and southern hakes. Though sources of exposure are complex because of multiple natural and anthropogenic PAH inputs, these results will facilitate the development of much needed health metrics for Gulf of Mexico benthos. Environ Toxicol Chem 2019;38:2740-2749. © 2019 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Growth and metabolism of adult polar cod (Boreogadus saida) in response to dietary crude oil.

The increasing human presence in the Arctic shelf seas, with the expansion of oil and gas industries and maritime shipping, poses a risk for Arctic marine organisms such as the key species polar cod (Boreogadus saida). The impact of dietary crude oil on growth and metabolism of polar cod was investigated in the early spring (March-April) when individuals are expected to be in a vulnerable physiological state with poor energy stores. Adult polar cod were exposed dietarily to three doses of Kobbe crude oil during an eight weeks period and followed by two weeks of depuration. Significant dose-responses in exposure biomarkers (hepatic ethoxyresorufine-O-deethylase [EROD] activity and 1-OH phenanthrene metabolites in bile) indicated that polycyclic aromatic hydrocarbons (PAHs) were bioavailable. Condition indices (i.e. Fulton's condition factor, hepatosomatic index), growth, whole body respiration, and total lipid content in the liver were monitored over the course of the experiment. The majority of females were immature, while a few had spawned during the season and showed low hepatic lipid content during the experiment. In contrast, males were all, except for one immature individual, in a post-spawning stage and had larger hepatic energy stores than females. Most specimens, independent of sex, showed a loss in weight, that was exacerbated by exposure to crude oil and low hepatic liver lipids. Furthermore, females exposed to crude oil showed a significant elevation of oxygen consumption compared to controls, although not dose-dependent. This study highlights the importance of the energy status of individuals for their response to a crude oil exposure.

Effects of dietary crude oil exposure on molecular and physiological parameters related to lipid homeostasis in polar cod (Boreogadus saida).

Polar cod is an abundant Arctic key species, inhabiting an ecosystem that is subjected to rapid climate change and increased petroleum related activities. Few studies have investigated biological effects of crude oil on lipid metabolism in this species, despite lipids being a crucial compound for Arctic species to adapt to the high seasonality in food abundance in their habitat. This study examines the effects of dietary crude oil exposure on transcription levels of genes related to lipid metabolism (peroxisome proliferator-activated receptors [ppar-α, ppar-γ], retinoic X receptor [rxr-ß], palmitoyl-CoA oxidase [aox1], cytochrome P4507A1 [cyp7α1]), reproduction (vitellogenin [vtg-ß], gonad aromatase [cyp19a1]) and biotransformation (cytochrome P4501A1 [cyp1a1], aryl hydrocarbon receptor [ahr2]). Exposure effects were also examined through plasma chemistry parameters. Additional fish were exposed to a PPAR-α agonist (WY-14,643) to investigate the role of PPAR-α in their lipid metabolism. The dose-dependent up-regulation of cyp1a1 reflected the activation of genes related to PAH biotransformation upon crude oil exposure. The crude oil exposure did not significantly alter the mRNA expression of genes involved in lipid homeostasis except for cyp7α1 transcription levels. Plasma levels of cholesterol and alanine transaminase showed significant alterations in fish exposed to crude oil at the end of the experiment. WY exposure induced a down-regulation of ppar-α, an effect contrary to studies performed on other fish species. In conclusion, this study showed clear effects of dietary crude oil exposure at environmentally relevant concentrations on xenobiotic biotransformation but revealed only weak alterations in the lipid metabolism of polar cod.

Induction of gonadal maturation at different temperatures in burbot Lota lota.

A rearing experiment was conducted to test whether temperature protocols that differed from a simulation of natural conditions might induce maturation after isothermal grow-out in burbot Lota lota. Lota lota were acclimated to two different temperature regimes: low temperature (LT), close to natural temperature at 4·0° C and elevated, high temperature (HT) at 8·5° C over 40 and 27 days respectively, with all fish then wintered for 47 days. Every second fish was treated with a gonadotropin-releasing hormone analogue. Maturational competence of oocytes was assessed with a germinal vesicle breakdown assay using a novel staining strategy. In both treatments, puberty and maturational progress occurred, characterised by an elevated gonado-somatic index and advanced gonadal stages (histological analysis). Progress of gonadal maturation was reflected by elevated plasma concentrations of testosterone and 11-ketosterone in males and 17ß-oestradiol in females. Ovulation was not observed. Sperm could be activated equally across treatments. In general, LT was more effective than HT treatment, indicated by advanced gonadal stages, higher numbers of oocytes undergoing germinal vesicle breakdown in vitro and elevated sex steroid levels. Hormone treatment could improve effectiveness at HT. In conclusion, less drastic temperature regimes as previously reported in combination with hormone treatments seem sufficient to induce maturation in L. lota after isothermal grow-out.

Early life stages of an arctic keystone species (Boreogadus saida) show high sensitivity to a water-soluble fraction of crude oil.

Increasing anthropogenic activities in the Arctic represent an enhanced threat for oil pollution in a marine environment that is already at risk from climate warming. In particular, this applies to species with free-living pelagic larvae that aggregate in surface waters and under the sea ice where hydrocarbons are likely to remain for extended periods of time due to low temperatures. We exposed the positively buoyant eggs of polar cod (Boreogadus saida), an arctic keystone species, to realistic concentrations of a crude oil water-soluble fraction (WSF), mimicking exposure of eggs aggregating under the ice to oil WSF leaking from brine channels following encapsulation in ice. Total hydrocarbon and polycyclic aromatic hydrocarbon levels were in the ng/L range, with most exposure concentrations below the limits of detection throughout the experiment for all treatments. The proportion of viable, free-swimming larvae decreased significantly with dose and showed increases in the incidence and severity of spine curvature, yolk sac alterations and a reduction in spine length. These effects are expected to compromise the motility, feeding capacity, and predator avoidance during critical early life stages for this important species. Our results imply that the viability and fitness of polar cod early life stages is significantly reduced when exposed to extremely low and environmentally realistic levels of aqueous hydrocarbons, which may have important implications for arctic food web dynamics and ecosystem functioning.

The role of molecular size in antioxidant activity of peptide fractions from Pacific hake (Merluccius productus) hydrolysates.

The antioxidative properties of Pacific hake hydrolysates and their peptidic fractions varying in molecular size were assessed. Hydrolysates produced by different proteases (Alcalase, bromelain, Flavourzyme, Protamex, Protease A"Amano"2, Protease N"Amano"K, Protin SD NY10, Umamizyme-K, Validase BNP-L, Validase FPexo) generally possessed good metal ion chelating (33-73% at 3mg/ml), DPPH radical scavenging (18-30% at 1mg/ml), ferric ion reducing power (abs700nm 0.36-0.86 at 3 mg/ml) and ABTS radical scavenging (47-85% at 0.067 mg/ml) activity, as well as a good capability to suppress lipid peroxidation in a linoleic acid model system. Peptide size (<1.4 kDa) was important for ABTS radical scavenging activity, whereas specific peptide composition (which depended on the particular protease used) was the governing factor for effective lipid peroxidation. Validase BNP-L was the most promising enzyme for producing Pacific hake hydrolysates with good antioxidative activity in various assays and similar effectiveness as the synthetic antioxidant BHT to inhibit lipid peroxidation.

Timing of the maturation transition in haddock Melanogrammus aeglefinus.

The timing of maturation in haddock Melanogrammus aeglefinus was examined using changes in gonad development, follicle stimulating hormone ß (FSH-ß) transcript expression profile, growth and condition of 1 year old females held under a common environment between the summer and winter solstices. The circumnuclear ring, cortical alveolus and vitellogenic oocyte stages were first observed in August, October and November, respectively. FSH-ß transcript levels did not change significantly until September but increased markedly thereafter in maturing fish. A combined analysis of the mean oocyte diameter of the leading cohort, histological staging and FSH-ß transcript profile provided evidence of a commitment to maturation by October or November. Contrary to that previously proposed for gadoid species, histological analysis of field-caught immature M. aeglefinus during the spawning season indicated that cortical alveolar, rather than circumnuclear ring, stage oocytes provided definitive evidence of maturation. A decrease in relative liver size following the summer solstice suggested a possible link between energy status and maturation.

Age and growth of the Argentine hake Merluccius hubbsi Marini, 1933 in the Brazilian South-Southeast Region during 1996-2001

This is the first detailed study on the age and growth of the Argentine hake Merluccius hubbsi in Brazilian waters. A total of 3,651 specimens obtained from surveys and sampling landings using trawlers and long-lines in the South-Southeast Region (21ºS to 34ºS) between 1996 and 2001 were analyzed. Readings of sliced and burned otoliths were conducted on 686 individuals (397 females, 129 males and 160 young - sex not identified), identifying opaque and translucent zones. Marginal increment and edge type percentage methods were used for validation, resulting in one ring per year, formed during summer-autumn. Fish with no more than eight rings and six years of age were found, the two first rings after the core being formed during the first year. Back-calculation methods were applied to length/age data and the parameters of von Bertalanffy's growth model were fitted based on average length per ring. The value of maximum theoretical length varied between 470.27 mm and 807.40 mm; growth coefficient varied from 0.1657 to 0.3555 year-1 and the theoretical age at length zero between -1.2846 and -0.4552 years. Growth of females and males differed significantly. The techniques applied and the results obtained are discussed and compared with studies on hake conducted in Uruguay and Argentina, providing important information for the management of the species in Brazil, where it presents clear signs of overexploitation

Este é o primeiro estudo detalhado sobre a idade e o crescimento da merluza Merluccius hubbsi no Brasil. Foram analisados 3.651 espécimes obtidos em cruzeiros de pesquisa e provenientes das frotas de arrasto e espinhel de fundo da região Sudeste-Sul do Brasil (21ºS a 34ºS) entre 1996 e 2001. Leituras de secções transversais de otólitos (corte, polimento e queima) foram realizadas para 686 indivíduos (397 fêmeas, 129 machos e 160 jovens com sexo não identificado), identificando-se zonas opacas e translúcidas. Foram calculados o incremento marginal e a porcentagem do tipo de borda, no tempo, para validação, resultando em um anel por ano, formado durante o verão-outono. Foram obtidos peixes com até oito anéis e seis anos de idade, sendo que os dois anéis depois do núcleo se formam durante o primeiro ano de vida. Foram aplicados métodos de retrocálculo aos dados de comprimento na idade e os parâmetros do modelo de von Bertalanffy foram estimados. O valor do comprimento máximo teórico variou entre 470,27 mm e 807,40 mm; a taxa de crescimento variou de 0,1657 a 0,3555 ano-1 e a idade teórica de comprimento zero entre -1,2846 e -0,4552 anos. O crescimento de fêmeas e machos difere significativamente. As técnicas aplicadas e os resultados obtidos foram discutidos, sendo comparados com estudos realizados no Uruguai e na Argentina e provendo informação importante para a gestão da espécie no Brasil, onde a merluza apresenta claros sinais de sobrepesca