Publications by authors named "Ørjan Karlsen"

23 Publications

  • Page 1 of 1

RADSex: A computational workflow to study sex determination using restriction site-associated DNA sequencing data.

Mol Ecol Resour 2021 Jul 9;21(5):1715-1731. Epub 2021 Mar 9.

Physiological Chemistry, Biocenter, University of Wuerzburg, Wuerzburg, Germany.

The study of sex determination and sex chromosome organization in nonmodel species has long been technically challenging, but new sequencing methodologies now enable precise and high-throughput identification of sex-specific genomic sequences. In particular, restriction site-associated DNA sequencing (RAD-Seq) is being extensively applied to explore sex determination systems in many plant and animal species. However, software specifically designed to search for and visualize sex-biased markers using RAD-Seq data is lacking. Here, we present RADSex, a computational analysis workflow designed to study the genetic basis of sex determination using RAD-Seq data. RADSex is simple to use, requires few computational resources, makes no prior assumptions about the type of sex-determination system or structure of the sex locus, and offers convenient visualization through a dedicated R package. To demonstrate the functionality of RADSex, we re-analysed a published data set of Japanese medaka, Oryzias latipes, where we uncovered a previously unknown Y chromosome polymorphism. We then used RADSex to analyse new RAD-Seq data sets from 15 fish species spanning multiple taxonomic orders. We identified the sex determination system and sex-specific markers in six of these species, five of which had no known sex-markers prior to this study. We show that RADSex greatly facilitates the study of sex determination systems in nonmodel species thanks to its speed of analyses, low resource usage, ease of application and visualization options. Furthermore, our analysis of new data sets from 15 species provides new insights on sex determination in fish.
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http://dx.doi.org/10.1111/1755-0998.13360DOI Listing
July 2021

Untangling mechanisms of crude oil toxicity: Linking gene expression, morphology and PAHs at two developmental stages in a cold-water fish.

Sci Total Environ 2021 Feb 2;757:143896. Epub 2020 Dec 2.

Institute of Marine Research, Bergen, Norway.

Early life stages of fish are highly sensitive to crude oil exposure and thus, short term exposures during critical developmental periods could have detrimental consequences for juvenile survival. Here we administered crude oil to Atlantic haddock (Melanogrammus aeglefinus) in short term (3-day) exposures at two developmental time periods: before first heartbeat, from gastrulation to cardiac cone stage (early), and from first heartbeat to one day before hatching (late). A frequent sampling regime enabled us to determine immediate PAH uptake, metabolite formation and gene expression changes. In general, the embryotoxic consequences of an oil exposure were more severe in the early exposure animals. Oil droplets on the eggshell resulted in severe cardiac and craniofacial abnormalities in the highest treatments. Gene expression changes of Cytochrome 1 a, b, c and d (cyp1a, b, c, d), Bone morphogenetic protein 10 (bmp10), ABC transporter b1 (abcb1) and Rh-associated G-protein (rhag) were linked to PAH uptake, occurrence of metabolites of phenanthrene and developmental and functional abnormalities. We detected circulation-independent, oil-induced gene expression changes and separated phenotypes linked to proliferation, growth and disruption of formation events at early and late developmental stages. Changes in bmp10 expression suggest a direct oil-induced effect on calcium homeostasis. Localized expression of rhag propose an impact on osmoregulation. Severe eye abnormalities were linked to possible inappropriate overexpression of cyp1b in the eyes. This study gives an increased knowledge about developmentally dependent effects of crude oil toxicity. Thus, our findings provide more knowledge and detail to new and several existing adverse outcome pathways of crude oil toxicity.
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http://dx.doi.org/10.1016/j.scitotenv.2020.143896DOI Listing
February 2021

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

PLoS One 2020 22;15(10):e0240307. Epub 2020 Oct 22.

Institute of Marine Research, Bergen, Norway.

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.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0240307PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580938PMC
December 2020

Effects of laboratory salmon louse infection on osmoregulation, growth and survival in Atlantic salmon.

Conserv Physiol 2020 26;8(1):coaa023. Epub 2020 Mar 26.

Reproduction and Developmental Biology, Institute of Marine Research (IMR), PO Box 1870, Nordnes, 5817 Bergen, Norway.

Anadromous Atlantic salmon () rely on long ocean migrations to build energy stores for maturation and spawning. In seawater, wild Atlantic salmon are threatened by high salmon lice () infestation levels resulting from intensive salmonid sea-cage aquaculture. Salmon lice infection can cause a stress response and an osmotic imbalance in the host. The lice infection intensity threshold values for these responses, however, remain to be identified in Atlantic salmon. In order to define this under laboratory conditions, individually tagged F1 wild origin Atlantic post-smolts (40 g) were infected with salmon lice copepodids or left as uninfected controls. Twenty-eight days post infection, infected post-smolts had a mean of 0.38 (range of 0.07-0.9) mobile lice g fish weight. During this period, specific growth rates (SGRs) were lower in infected than control fish (0.4 vs 1.0% day). Higher plasma Na, Cl and osmolality in infected fish also indicate osmoregulatory impairment. SGR correlated negatively with plasma Na, Cl, osmolality and cortisol in the infected, but not in the control group. Infection intensity (lice g fish) correlated positively with mortality rate and plasma Na, Cl, osmolality and cortisol and correlated negatively with SGR and condition factor. Calculated lice intensity threshold values for changes in plasma ions were 0.18 lice g for plasma Cl, and 0.22 lice g for plasma Na. Moribund infected fish occurred at infection intensities above 0.2 lice g, and these fish had extreme plasma Cl, Na, osmolality and cortisol levels. There was a positive correlation between plasma cortisol and plasma Na, Cl and osmolality in infected fish. This study provides vital information that can be used to define thresholds in the monitoring and conservation of wild Atlantic salmon populations affected by aquaculture-driven salmon lice infestations.
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http://dx.doi.org/10.1093/conphys/coaa023DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7098368PMC
March 2020

Evaluation of a national operational salmon lice monitoring system-From physics to fish.

PLoS One 2018 31;13(7):e0201338. Epub 2018 Jul 31.

Institute of Marine Research, Bergen, Norway.

The Norwegian government has decided that the aquaculture industry shall grow, provided that the growth is environmentally sustainable. Sustainability is scored based on the mortality of wild salmonids caused by the parasitic salmon lice. Salmon lice infestation pressure has traditionally been monitored through catching wild sea trout and Arctic char using nets or traps or by trawling after Atlantic salmon postsmolts. However, due to that the Norwegian mainland coastline is nearly 25 000 km, complementary methods that may be used in order to give complete results are needed. We have therefore developed an operational salmon lice model, which calculates the infestation pressure all along the coast in near real-time based on a hydrodynamical ocean model and a salmon lice particle tracking model. The hydrodynamic model generally shows a negative temperature bias and a positive salinity bias compared to observations. The modeled salmon lice dispersion correlates with measured lice on wild salmonids caught using traps or nets. This allows for using two complementary data sources in order to determine the infestation pressure of lice originating from fish farms on wild salmonids, and thereby provide an improved monitoring system for assessing risk and sustainability which forms the basis for knowledge-based advice to management authorities.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0201338PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6067748PMC
January 2019

Oil droplet fouling and differential toxicokinetics of polycyclic aromatic hydrocarbons in embryos of Atlantic haddock and cod.

PLoS One 2017 5;12(7):e0180048. Epub 2017 Jul 5.

Institute of Marine Research, Bergen, Norway.

The impact of crude oil pollution on early life stages (ELS) of fish, including larvae and embryos, has received considerable attention in recent years. Of the organic components present in crude oil, polycyclic aromatic hydrocarbons (PAHs) are considered the main class of compounds responsible for toxic effects in marine organisms. Although evidence suggests that they are more toxic, alkylated PAHs remain much less studied than their unsubstituted congeners. Recently, it was established that embryos of Atlantic haddock (Melanogrammus aeglefinus) are particularly sensitive to dispersed crude oil, and it was hypothesized that this was caused by direct interaction with crude oil droplets, which adhered to the chorion of exposed embryos. Such a phenomenon would increase the potential for uptake of less water-soluble compounds, including alkylated PAHs. In the current study, we compared the uptake of parent and alkylated PAHs in Atlantic cod (Gadus morhua) and haddock embryos exposed to dispersed crude oil at a range of environmentally relevant concentrations (10-600 μg oil/liter seawater). Although the species are biologically very similar, the cod chorion does not become fouled with oil droplets, even when the two species are exposed to dispersions of crude oil droplets under similar conditions. A close correlation between the degree of fouling and toxicological response (heart defects, craniofacial malformation) was observed. Oil droplet fouling in haddock led to both quantitative and qualitative differences in PAH uptake. Finally, kinetic data on a large suite of PAHs showed differential elimination, suggesting differential metabolism of unsubstituted versus alkylated compounds.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0180048PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5497984PMC
September 2017

Crude oil exposures reveal roles for intracellular calcium cycling in haddock craniofacial and cardiac development.

Sci Rep 2016 08 10;6:31058. Epub 2016 Aug 10.

Institute of Marine Research, P.O. Box 1870, Nordnes, NO-5817, Bergen, Norway.

Recent studies have shown that crude oil exposure affects cardiac development in fish by disrupting excitation-contraction (EC) coupling. We previously found that eggs of Atlantic haddock (Melanogrammus aeglefinus) bind dispersed oil droplets, potentially leading to more profound toxic effects from uptake of polycyclic aromatic hydrocarbons (PAHs). Using lower concentrations of dispersed crude oil (0.7-7 μg/L ∑PAH), here we exposed a broader range of developmental stages over both short and prolonged durations. We quantified effects on cardiac function and morphogenesis, characterized novel craniofacial defects, and examined the expression of genes encoding potential targets underlying cardiac and craniofacial defects. Because of oil droplet binding, a 24-hr exposure was sufficient to create severe cardiac and craniofacial abnormalities. The specific nature of the craniofacial abnormalities suggests that crude oil may target common craniofacial and cardiac precursor cells either directly or indirectly by affecting ion channels and intracellular calcium in particular. Furthermore, down-regulation of genes encoding specific components of the EC coupling machinery suggests that crude oil disrupts excitation-transcription coupling or normal feedback regulation of ion channels blocked by PAHs. These data support a unifying hypothesis whereby depletion of intracellular calcium pools by crude oil-derived PAHs disrupts several pathways critical for organogenesis in fish.
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http://dx.doi.org/10.1038/srep31058DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4979050PMC
August 2016

The two-step development of a duplex retina involves distinct events of cone and rod neurogenesis and differentiation.

Dev Biol 2016 08 30;416(2):389-401. Epub 2016 Jun 30.

Department of Biology, University of Bergen, NO-5020 Bergen, Norway.

Unlike in mammals, persistent postembryonic retinal growth is a characteristic feature of fish, which includes major remodeling events that affect all cell types including photoreceptors. Consequently, visual capabilities change during development, where retinal sensitivity to different wavelengths of light (photopic vision), -and to limited photons (scotopic vision) are central capabilities for survival. Differently from well-established model fish, Atlantic cod has a prolonged larval stage where only cone photoreceptors are present. Rods do not appear until juvenile transition (metamorphosis), a hallmark of indirect developing species. Previously we showed that whole gene families of lws (red-sensitive) and sws1 (UV-sensitive) opsins have been lost in cod, while rh2a (green-sensitive) and sws2 (blue-sensitive) genes have tandem duplicated. Here, we provide a comprehensive characterization of a two-step developing duplex retina in Atlantic cod. The study focuses on cone subtype dynamics and delayed rod neurogenesis and differentiation in all cod life stages. Using transcriptomic and histological approaches we show that different opsins disappear in a topographic manner during development where central to peripheral retina is a key axis of expressional change. Early cone differentiation was initiated in dorso-temporal retina different from previously described in fish. Rods first appeared during initiation of metamorphosis and expression of the nuclear receptor transcription factor nr2e3-1, suggest involvement in rod specification. The indirect developmental strategy thus allows for separate studies of cones and rods development, which in nature correlates with visual changes linked to habitat shifts. The clustering of key retinal genes according to life stage, suggests that Atlantic cod with its sequenced genome may be an important resource for identification of underlying factors required for development and function of photopic and scotopic vision.
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http://dx.doi.org/10.1016/j.ydbio.2016.06.041DOI Listing
August 2016

The Ontogeny and Brain Distribution Dynamics of the Appetite Regulators NPY, CART and pOX in Larval Atlantic Cod (Gadus morhua L.).

PLoS One 2016 21;11(4):e0153743. Epub 2016 Apr 21.

Department of Biology, University of Bergen, Bergen, Norway.

Similar to many marine teleost species, Atlantic cod undergo remarkable physiological changes during the early life stages with concurrent and profound changes in feeding biology and ecology. In contrast to the digestive system, very little is known about the ontogeny and the localization of the centers that control appetite and feed ingestion in the developing brain of fish. We examined the expression patterns of three appetite regulating factors (orexigenic: neuropeptide Y, NPY; prepro-orexin, pOX and anorexigenic: cocaine- and amphetamine-regulated transcript, CART) in discrete brain regions of developing Atlantic cod using chromogenic and double fluorescent in situ hybridization. Differential temporal and spatial expression patterns for each appetite regulator were found from first feeding (4 days post hatch; dph) to juvenile stage (76 dph). Neurons expressing NPY mRNA were detected in the telencephalon (highest expression), diencephalon, and optic tectum from 4 dph onward. CART mRNA expression had a wider distribution along the anterior-posterior brain axis, including both telencephalon and diencephalon from 4 dph. From 46 dph, CART transcripts were also detected in the olfactory bulb, region of the nucleus of medial longitudinal fascicle, optic tectum and midbrain tegmentum. At 4 and 20 dph, pOX mRNA expression was exclusively found in the preoptic region, but extended to the hypothalamus at 46 and 76 dph. Co-expression of both CART and pOX genes were also observed in several hypothalamic neurons throughout larval development. Our results show that both orexigenic and anorexigenic factors are present in the telencephalon, diencephalon and mesencephalon in cod larvae. The telencephalon mostly contains key factors of hunger control (NPY), while the diencephalon, and particularly the hypothalamus may have a more complex role in modulating the multifunctional control of appetite in this species. As the larvae develop, the overall progression in temporal and spatial complexity of NPY, CART and pOX mRNAs expression might be correlated to the maturation of appetite control regulation. These observations suggest that teleost larvae continue to develop the regulatory networks underlying appetite control after onset of exogenous feeding.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0153743PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4839749PMC
September 2016

First feed affects the expressions of microRNA and their targets in Atlantic cod.

Br J Nutr 2016 Apr 9;115(7):1145-54. Epub 2016 Feb 9.

1Faculty of Biosciences and Aquaculture,Nord University,Postbox 1490,8049 Bodø,Norway.

To our knowledge, there is no report on microRNA (miRNA) expression and their target analysis in relation to the type of the first feed and its effect on the further growth of fish. Atlantic cod (Gadus morhua) larvae have better growth and development performance when fed natural zooplankton as a start-feed, as compared with those fed typical aquaculture start-feeds. In our experiment, two groups of Atlantic cod larvae were fed reference feed (zooplankton, mostly copepods, filtered from a seawater pond) v. aquaculture feeds: enriched rotifers (Brachionus sp.) and later brine shrimp (Artemia salina). We examined the miRNA expressions of six defined developmental stages as determined and standardised by body length from first feeding for both diet groups. We found eight miRNA (miR-9, miR-19a, miR-130b, miR-146, miR-181a, miR-192, miR-206 and miR-11240) differentially expressed between the two feeding groups in at least one developmental stage. We verified the next-generation sequencing data using real-time RT-PCR. We found 397 putative targets (mRNA) to the differentially expressed miRNA; eighteen of these mRNA showed differential expression in at least one stage. The patterns of differentially expressed miRNA and their putative target mRNA were mostly inverse, but sometimes also concurrent. The predicted miRNA targets were involved in different pathways, including metabolic, phototransduction and signalling pathways. The results of this study provide new nutrigenomic information on the potential role of miRNA in mediating nutritional effects on growth during the start-feeding period in fish larvae.
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http://dx.doi.org/10.1017/S0007114516000155DOI Listing
April 2016

1H NMR metabolic profiling of cod (Gadus morhua) larvae: potential effects of temperature and diet composition during early developmental stages.

Biol Open 2015 Nov 6;4(12):1671-8. Epub 2015 Nov 6.

National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029, Bergen N-5817, Norway.

Marine aquaculture offers a great source of protein for the increasing human population, and farming of, for example, Atlantic salmon is a global industry. Atlantic cod farming however, is an example of a promising industry where the potential is not yet realized. Research has revealed that a major bottleneck to successful farming of cod is poor quality of the larvae and juveniles. A large research program was designed to increase our understanding of how environmental factors such as temperature and nutrition affects cod larvae development. Data on larvae growth and development were used together with nuclear magnetic resonance. The NMR data indicated that the temperature influenced the metabolome of the larvae; differences were related to osmolytes such as betaine/TMAO, the amino acid taurine, and creatine and lactate which reflect muscle activity. The larvae were fed Artemia from stage 2, and this was probably reflected in a high taurine content of older larvae. Larvae fed with copepods in the nutrition experiment also displayed a high taurine content, together with higher creatine and betaine/TMAO content. Data on the cod larvae metabolome should be coupled to data on gene expression, in order to identify events which are regulated on the genetic level versus regulation resulting from temperature or nutrition during development, to fully understand how the environment affects larval development.
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http://dx.doi.org/10.1242/bio.014431DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4736036PMC
November 2015

Diet affects the redox system in developing Atlantic cod (Gadus morhua) larvae.

Redox Biol 2015 Aug 12;5:308-318. Epub 2015 Jun 12.

National Institute of Nutrition and Seafood Research (NIFES), PO Box 2029, NO-5817 Bergen, Norway. Electronic address:

The growth and development of marine fish larvae fed copepods is superior to those fed rotifers, but the underlying molecular reasons for this are unclear. In the following study we compared the effects of such diets on redox regulation pathways during development of Atlantic cod (Gadus morhua) larvae. Cod larvae were fed a control diet of copepods or the typical rotifer/Artemia diet commonly used in commercial marine fish hatcheries, from first feeding until after metamorphosis. The oxidised and reduced glutathione levels, the redox potential, and the mRNA expression of 100 genes in redox system pathways were then compared between treatments during larval development. We found that rotifer/Artemia-fed cod larvae had lower levels of oxidised glutathione, a more reduced redox potential, and altered expression of approximately half of the redox system genes when compared to copepod-fed larvae. This rotifer/Artemia diet-induced differential regulation of the redox system was greatest during periods of suboptimal growth. Upregulation of the oxidative stress response transcription factor, nrf2, and NRF2 target genes in rotifer/Artemia fed larvae suggest this diet induced an NRF2-mediated oxidative stress response. Overall, the data demonstrate that nutritional intake plays a role in regulating the redox system in developing fish larvae. This may be a factor in dietary-induced differences observed in larval growth.
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http://dx.doi.org/10.1016/j.redox.2015.06.003DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4488531PMC
August 2015

Copepods enhance nutritional status, growth and development in Atlantic cod (Gadus morhua L.) larvae - can we identify the underlying factors?

PeerJ 2015 19;3:e902. Epub 2015 May 19.

National Institute of Nutrition and Seafood Research , Nordnes, Bergen , Norway.

The current commercial production protocols for Atlantic cod depend on enriched rotifers and Artemia during first-feeding, but development and growth remain inferior to fish fed natural zooplankton. Two experiments were conducted in order to identify the underlying factors for this phenomenon. In the first experiment (Exp-1), groups of cod larvae were fed either (a) natural zooplankton, mainly copepods, increasing the size of prey as the larvae grew or (b) enriched rotifers followed by Artemia (the intensive group). In the second experiment (Exp-2), two groups of larvae were fed as in Exp-1, while a third group was fed copepod nauplii (approximately the size of rotifers) throughout the larval stage. In both experiments, growth was not significantly different between the groups during the first three weeks after hatching, but from the last part of the rotifer feeding period and onwards, the growth of the larvae fed copepods was higher than that of the intensive group. In Exp-2, the growth was similar between the two copepod groups during the expeimental period, indicating that nutrient composition, not prey size caused the better growth on copepods. Analyses of the prey showed that total fatty acid composition and the ratio of phospholipids to total lipids was slightly different in the prey organisms, and that protein, taurine, astaxanthin and zinc were lower on a dry weight basis in rotifers than in copepods. Other measured nutrients as DHA, all analysed vitamins, manganese, copper and selenium were similar or higher in the rotifers. When compared to the present knowledge on nutrient requirements, protein and taurine appeared to be the most likely limiting nutrients for growth in cod larvae fed rotifers and Artemia. Larvae fed rotifers/Artemia had a higher whole body lipid content than larvae fed copepods at the end of the experiment (stage 5) after the fish had been fed the same formulated diet for approximately 2 weeks.
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http://dx.doi.org/10.7717/peerj.902DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4451035PMC
June 2015

Unexpected interaction with dispersed crude oil droplets drives severe toxicity in Atlantic haddock embryos.

PLoS One 2015 29;10(4):e0124376. Epub 2015 Apr 29.

Institute of Marine Research, Nordnes, Bergen, Norway.

The toxicity resulting from exposure to oil droplets in marine fish embryos and larvae is still subject for debate. The most detailed studies have investigated the effects of water-dissolved components of crude oil in water accommodated fractions (WAFs) that lack bulk oil droplets. Although exposure to dissolved petroleum compounds alone is sufficient to cause the characteristic developmental toxicity of crude oil, few studies have addressed whether physical interaction with oil micro-droplets are a relevant exposure pathway for open water marine speices. Here we used controlled delivery of mechanically dispersed crude oil to expose pelagic embryos and larvae of a marine teleost, the Atlantic haddock (Melanogrammus aeglefinus). Haddock embryos were exposed continuously to two different concentrations of dispersed crude oil, high and low, or in pulses. By 24 hours of exposure, micro-droplets of oil were observed adhering and accumulating on the chorion, accompanied by highly elevated levels of cyp1a, a biomarker for exposure to aromatic hydrocarbons. Embryos from all treatment groups showed abnormalities representative of crude oil cardiotoxicity at hatch (5 days of exposure), such as pericardial and yolk sac edema. Compared to other species, the frequency and severity of toxic effects was higher than expected for the waterborne PAH concentrations (e.g., 100% of larvae had edema at the low treatment). These findings suggest an enhanced tissue uptake of PAHs and/or other petroleum compounds from attached oil droplets. These studies highlight a novel property of haddock embryos that leads to greater than expected impact from dispersed crude oil. Given the very limited number of marine species tested in similar exposures, the likelihood of other species with similar properties could be high. This unanticipated result therefore has implications for assessing the ecological impacts of oil spills and the use of methods for dispersing oil in the open sea.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0124376PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4414579PMC
February 2016

Cortisol treatment of prespawning female cod affects cytogenesis related factors in eggs and embryos.

Gen Comp Endocrinol 2013 Aug 6;189:84-95. Epub 2013 May 6.

Institute of Marine Research, P. O. Box 1870, 5817 Bergen, Norway.

A stable supply of viable eggs and embryos is crucial for successful farming of Atlantic cod. Stress during broodstock rearing can have negative effects on offspring, but little is known about the molecular mechanisms that cause abnormal development. Maternally transferred mRNAs have been shown to be essential for normal development, and stress may therefore influence their expression and the subsequent embryonic development. We investigated if mimicked stress in cod females affects mRNA concentrations in eggs/embryos, and if this can be linked to viability of embryos. Three weeks before peak spawning, 20 fish were intraperitoneally implanted with either cortisol-containing or cortisol-free (sham) osmotic pumps. At peak spawning all individuals were stripped and eggs were fertilized and incubated until hatching. Samples were collected from unfertilized eggs and embryos for analysis of gene expression (microarray), viability, steroids and vitellogenin. Plasma concentration of cortisol (ng/ml) in treated females was significantly higher at spawning (127.1±20.9) than that of sham control (11.3±6.7). This difference was also reflected in eggs and embryos. Percent fertilization, asymmetric cell division and hatching were not affected. However, numerous genes were differentially expressed in eggs and embryos in response to elevated cortisol, especially in maternal (oocyte and blastula) stages. Among these differentially expressed genes, some were found to be linked to cytogenesis (stxbp6, fbxw2, capn12, thbs4, sytl2, coro1c, sel1l3), induction of mesodermal fate (fgfrl1) and import of the glucocorticoid receptor to the cell nucleus (ipo7). Gene ontology overrepresentation analysis on the whole set of differentially expressed genes at maternal stages (539 genes) revealed enriched activity in membrane associated regions, which largely corresponds to cytogenesis related processes. These results suggest that despite no visible phenotypic effects in early embryos, broodstock stress affects the egg/embryonic transcriptome, especially in relation to cytogenesis. Furthermore, effects related to egg/embryo phenotypes are difficult to measure at early stages of development, and instead might become apparent at later life stages.
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http://dx.doi.org/10.1016/j.ygcen.2013.04.028DOI Listing
August 2013

Pituitary gonadotropin and testicular gonadotropin receptor expression in Atlantic cod (Gadusmorhua L.) during the first reproductive season: Effects of photoperiod modulation.

Gen Comp Endocrinol 2011 Aug 14;173(1):111-9. Epub 2011 May 14.

Utrecht University, Faculty of Sciences, Department of Biology, Division Developmental Biology, P.O. Box 80058, 3508 TB Utrecht, The Netherlands.

Pituitary mRNA levels of gonadotropin β-subunits and of their cognate receptors in the testis were studied during puberty in Atlantic cod under normal and experimental photoperiod conditions that suppressed, delayed or accelerated testis maturation. Results are discussed in context with changes in testicular histology and plasma androgen levels, considered as end points of gonadotropic regulation. Up-regulation of fshb was closely associated with the onset of puberty, decreased when spermatogenesis was completed and reached minimum levels after spawning. These results demonstrate, for the first time using an experimental approach, that activation of Fsh-dependent signaling is associated with spermatogonial proliferation and formation of spermatogenic cysts. Changes in fshr expression were less prominent and could be explained by changes in the cellular composition and RNA content of cod testis tissue. At more advanced stages of development (spermiogenesis, spermiation and spawning), lhb and, one month later, lhcgr transcript levels increased and reached peak values in spawning fish, in a positive feedback loop involving plasma androgens and Lh/Lhcgr-dependent signaling. This loop was broken by a loss of lhb expression at the end of the spawning season. Continuous light (LL) from summer solstice, ~8 months prior to spawning, suppressed the start of testis maturation and the changes in gonadotropin and receptor mRNA levels, while LL from winter solstice initially up-regulated lhb and lhcgr expression, before resulting in a precocious termination of the spawning season and low expression of all four genes. Our studies provide experimental evidence for a clear functional discrimination of cod gonadotropins.
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http://dx.doi.org/10.1016/j.ygcen.2011.05.002DOI Listing
August 2011

Control of puberty in farmed fish.

Gen Comp Endocrinol 2010 Feb 13;165(3):483-515. Epub 2009 May 13.

Institute of Marine Research, PO Box 1870 Nordnes, 5817 Bergen, Norway.

Puberty comprises the transition from an immature juvenile to a mature adult state of the reproductive system, i.e. the individual becomes capable of reproducing sexually for the first time, which implies functional competence of the brain-pituitary-gonad (BPG) axis. Early puberty is a major problem in many farmed fish species due to negative effects on growth performance, flesh composition, external appearance, behaviour, health, welfare and survival, as well as possible genetic impact on wild populations. Late puberty can also be a problem for broodstock management in some species, while some species completely fail to enter puberty under farming conditions. Age and size at puberty varies between and within species and strains, and are modulated by genetic and environmental factors. Puberty onset is controlled by activation of the BPG axis, and a range of internal and external factors are hypothesised to stimulate and/or modulate this activation such as growth, adiposity, feed intake, photoperiod, temperature and social factors. For example, there is a positive correlation between rapid growth and early puberty in fish. Age at puberty can be controlled by selective breeding or control of photoperiod, feeding or temperature. Monosex stocks can exploit sex dimorphic growth patterns and sterility can be achieved by triploidisation. However, all these techniques have limitations under commercial farming conditions. Further knowledge is needed on both basic and applied aspects of puberty control to refine existing methods and to develop new methods that are efficient in terms of production and acceptable in terms of fish welfare and sustainability.
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http://dx.doi.org/10.1016/j.ygcen.2009.05.004DOI Listing
February 2010

Quantification of gonadotropin subunits GPalpha, FSHbeta, and LHbeta mRNA expression from Atlantic cod (Gadus morhua) throughout a reproductive cycle.

Comp Biochem Physiol B Biochem Mol Biol 2009 Jul 1;153(3):288-95. Epub 2009 Apr 1.

Institute of Marine Research Austevoll, N-5392 Storebø, Norway.

To elucidate the role of the gonadotropins in Atlantic cod (Gadus morhua), complete coding sequences with partially or fully un-translated regions for the three subunits GPalpha, FSHbeta, and LHbeta were determined. The sequences of the corresponding genomic loci were also determined, allowing the design of mRNA-targeting quantitative PCR assays. Relative expression was analyzed during a complete seasonal sexual maturation cycle in Atlantic cod females. Increasing levels of lhbeta mRNA were observed during gonadal growth, peaking at spawning in February-March which corresponds to maximum gonadosomatic index. In contrast, both gpalpha and fshbeta gradually increased to a peak in December, two months before spawning started, and decreased in January just prior to spawning. Both mRNAs increased again and remained high during the spawning season, with a decline at the end of the spawning period, a further decrease in spent females, followed by a new gradual increase concurrent with the start of the next reproductive cycle. In addition to its role in vitellogenesis prior to spawning, FSH seems to have additional functions during the spawning period, possibly related to vitellogenesis that runs in parallel with final oocyte maturation and ovulation of the multiple batch spawner Atlantic cod.
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http://dx.doi.org/10.1016/j.cbpb.2009.03.011DOI Listing
July 2009

Photoperiod-modulated testis maturation in Atlantic cod (Gadus morhua, L.).

Biol Reprod 2009 Apr 26;80(4):631-40. Epub 2008 Nov 26.

Faculty of Sciences, Department of Biology, Division of Endocrinology & Metabolism, Utrecht University, Utrecht, the Netherlands.

Precocious male puberty is a significant problem in Atlantic cod aquaculture. While photoperiod manipulation can inhibit testis growth, a detailed analysis of effects on spermatogenesis is missing. Starting July 1, 2004, prepubertal fish were exposed to different photoperiod regimens in indoor tanks for 17 mo. Testis histology, germ cell dynamics (proliferation and apoptosis), and plasma androgen levels were analyzed. In the natural light (NL) group, testis growth started in September 2004 and was completed in February 2005, when a 2-mo spawning period started. In the constant light (LL) group, none or very few spermatogenic cysts were recruited into spermatogenesis, and apoptotic germ cell loss was high. A change of photoperiod from NL to LL at winter solstice (December 21, 2004) resulted in premature (2 mo) completion of the reproductive cycle, while changing from LL to NL at winter solstice triggered faster than normal testis development. Plasma testosterone levels increased in the NL group from spermatogonial proliferation toward meiosis, while those of 11-ketotestosterone increased toward spermiogenesis and spermiation. Plasma androgen levels did not rise under LL conditions. Comparing fish with developing testes from all groups indicated that low androgen levels were associated with a high incidence of spermatogonial apoptosis; we also found that androgen receptor mRNA expression was most prominent in Sertoli cells in contact with growing spermatogonial clones. Our data show that an inhibitory photoperiod (LL) reduced or blocked differentiation of spermatogonia, increased apoptosis (particularly among proliferating spermatogonia), and was associated with reduced androgen levels, a situation possibly reflecting insufficient gonadotropic stimulation.
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http://dx.doi.org/10.1095/biolreprod.108.071340DOI Listing
April 2009

Gonadal development and associated changes in liver size and sexual steroids during the reproductive cycle of captive male and female Atlantic cod (Gadus morhua L.).

Comp Biochem Physiol A Mol Integr Physiol 2003 Nov;136(3):641-53

Department of Aquaculture, Institute of Marine Research, N-5392, Storebo, Norway.

Gametogenesis in female and male Atlantic cod (Gadus morhua L.) was investigated by sampling blood plasma and gonadal tissue from 19 to 33-month-old fish. The reproductive cycles of both female and male Atlantic cod are characterized by distinct annual variations in gonadal size and developmental stage and these are associated with changes in sex steroids and liver size. I(H) did not change during early gonadal development, but both spent females and males had lower I(H) than late maturing females and spermiating males, respectively. In females I(G) was correlated to plasma E2 levels and they were highest in spawning females. The lowest levels during the reproductive cycle were observed in spent females. Plasma T levels were low throughout ovarian development, and were at a minimum in spent females. 11-ketotestosterone in plasma of males increased rapidly during spermiation, while T increased at earlier testicular stages and reached maximum during spermiation. High plasma levels of steroids in male and female cod during spawning serve to promote further development and growth of less advanced stages of germ cells.
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http://dx.doi.org/10.1016/s1095-6433(03)00215-0DOI Listing
November 2003

Spermatogenesis and related plasma androgen levels in Atlantic halibut (Hippoglossus hippoglossus L.).

Comp Biochem Physiol A Mol Integr Physiol 2002 Jul;132(3):567-75

Institute of Marine Research, Austevoll Aquaculture Research Station, 5392 Storebø, Norway.

Spermatogenesis in male Atlantic halibut (Hippoglossus hippoglossus L.) was investigated by sampling blood plasma and testicular tissue from 15-39-month-old fish. The experiment covered a period in which all fish reached puberty and completed sexual maturation at least once. The germinal compartment in Atlantic halibut testis appears to be organized in branching lobules of the unrestricted spermatogonial type, because spermatocysts with spermatogonia were found throughout the testis. Spermatogenesis was characterized histologically, and staged according to the most advanced type of germ cell present: spermatogonia (Stage I), spermatogonia and spermatocytes (Stage II), spermatogonia, spermatocytes and spermatids (Stage III), spermatogonia, spermatocytes, spermatids and spermatozoa (Stage IV), and regressing testis (Stage V). Three phases could be distinguished: first, an initial phase with low levels of circulating testosterone (T; quantified by RIA) and 11-ketotestosterone (11-KT; quantified by ELISA), spermatogonial proliferation, and subsequently the initiation of meiosis marked by the formation of spermatocytes (Stage I and II). Secondly, a phase with increasing T and 11-KT levels and with haploid germ cells including spermatozoa present in the testis (Stage III and IV). Thirdly, a phase with low T and 11-KT levels and a regressing testis with Sertoli cells displaying signs of phagocytotic activity (Stage V). Circulating levels of 11-KT were at least four-fold higher than those of T during all stages of spermatogenesis. Increasing plasma levels of T and 11-KT were associated with increasing testicular mass throughout the reproductive cycle. The absolute level of, or the relation between, testis growth and circulating androgens were not significantly different in first time spawners compared to fish that underwent their second spawning season. These results provide reference levels for Atlantic halibut spermatogenesis.
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http://dx.doi.org/10.1016/s1095-6433(02)00092-2DOI Listing
July 2002