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

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

Dandelion polysaccharide suppresses lipid oxidation in Antarctic krill (Euphausia superba).

Antarctic krill (Euphausia superba) are rich in unsaturated lipids that are susceptible to oxidation even under frozen storage. In this study, Antarctic krill was treated with dandelion polysaccharide (DP) and stored frozen at -8 °C for 12 months. The changes in thiobarbituric acid reactants (TBARs), peroxide values (PVs), free fatty acid (FFA) content, fatty acid composition and sensory quality score were evaluated. Treatment with DP maintained the PV, TBAR and FFA contents and sensory quality score and increased the unsaturated fatty acid contents. Results showed that DP treatment could be a promising way to suppresses lipid oxidation and extend the shelf life of Antarctic krill.

Effects of chronic crude oil exposure on early developmental stages of the Northern krill (Meganyctiphanes norvegica).

Rising oil and gas activities in northern high latitudes have led to an increased risk of petroleum pollution in these ecosystems. Further, seasonal high UV radiation at high latitudes may elevate photo-enhanced toxicity of petroleum pollution to marine organisms. Zooplanktons are a key ecological component of northern ecosystems; therefore, it is important to assess their sensitivity to potential pollutants of oil and gas activity. As ontogenetic development may be particularly sensitive, the aim of this study was to examine the impact of chronic exposure to oil water dispersion (OWD) on development and feeding of early life stages of the Northern krill, Meganyctiphanes norvegica. In a range of experiments, embryonic, nonfeeding, and feeding larval stages were exposed to concentrations of between 0.01 and 0.1 mg/L of oil or photo-modified oil for 19 and 21 d. No significant effects on egg respiration, hatching success, development, length and larval survival were observed from these treatments. Similarly, evolution of fatty acid composition patterns during ontogenetic development was unaffected. The results indicates a high degree of resilience of these early developmental stages to such types and concentrations of pollutants. However, feeding and motility in later calyptopis-stage larvae were significantly impaired at exposure of 0.1 mg/L oil. Data indicate that feeding larval stage of krill was more sensitive to OWD than early nonfeeding life stages. This might be attributed to the narcotic effects of oil pollutants, their direct ingestion, or accumulated adverse effects over early development.

KrillDB: A de novo transcriptome database for the Antarctic krill (Euphausia superba).

Antarctic krill (Euphausia superba) is a key species in the Southern Ocean with an estimated biomass between 100 and 500 million tonnes. Changes in krill population viability would have catastrophic effect on the Antarctic ecosystem. One looming threat due to elevated levels of anthropogenic atmospheric carbon dioxide (CO2) is ocean acidification (lowering of sea water pH by CO2 dissolving into the oceans). The genetics of Antarctic krill has long been of scientific interest for both for the analysis of population structure and analysis of functional genetics. However, the genetic resources available for the species are relatively modest. We have developed the most advanced genetic database on Euphausia superba, KrillDB, which includes comprehensive data sets of former and present transcriptome projects. In particular, we have built a de novo transcriptome assembly using more than 360 million Illumina sequence reads generated from larval krill including individuals subjected to different CO2 levels. The database gives access to: 1) the full list of assembled genes and transcripts; 2) their level of similarity to transcripts and proteins from other species; 3) the predicted protein domains contained within each transcript; 4) their predicted GO terms; 5) the level of expression of each transcript in the different larval stages and CO2 treatments. All references to external entities (sequences, domains, GO terms) are equipped with a link to the appropriate source database. Moreover, the software implements a full-text search engine that makes it possible to submit free-form queries. KrillDB represents the first large-scale attempt at classifying and annotating the full krill transcriptome. For this reason, we believe it will constitute a cornerstone of future approaches devoted to physiological and molecular study of this key species in the Southern Ocean food web.

Dietary exposure of Antarctic krill to p,p'-DDE: uptake kinetics and toxicological sensitivity in a key polar species.

This study evaluated the dietary uptake kinetics and sublethal toxicity of p,p'-dichlorodiphenyl dichloroethylene (p,p'-DDE) in Antarctic krill. The uptake rate constant (characterised by the seawater volume stripped of contaminant sorbed to algae) of 200 ± 0.32 mL g(-1) wet weight h(-1), average absorption efficiency of 86 ± 13% and very low elimination rate constant of 5 × 10(-6) ± 0.0031 h(-1) demonstrate the importance of feeding for p,p'-DDE bioaccumulation in Antarctic krill. Faecal egestion of unabsorbed p,p'-DDE of 8.1 ± 2.7% indicates that this pathway contributes considerably to p,p'-DDE sinking fluxes. A median internal effective concentration (IEC50) of 15 mmol/kg lipid weight for complete immobility indicates baseline toxicity and that Antarctic krill exhibit comparable toxicological sensitivity as temperate species under similar 10 d exposure conditions. These findings support the critical body residue approach and provide insight to the role of Antarctic krill in the biogeochemical cycling of p,p'-DDE in the Southern Ocean.

Behavioural sensitivity of a key Southern Ocean species (Antarctic krill, Euphausia superba) to p,p'-DDE exposure.

Persistent organic pollutants (POPs) have been frequently measured throughout the Southern Ocean food web for which little information is available to assess the potential risks of POP exposure. The current study evaluated the toxicological sensitivity of a key Southern Ocean species, Antarctic krill, to aqueous exposure of p,p'-dichlorodiphenyl dichloroethylene (p,p'-DDE). Behavioural endpoints were used as indicators of sublethal toxicity. Immediate behavioural responses (partial immobility and tail flicking) most likely reflect neurotoxicity, while the p,p'-DDE body residue causing a median level of sublethal toxicity in Antarctic krill following 96h exposure (IEC50(sublethal toxicity)=3.9±0.21mmol/kg lipid weight) is comparable to those known to cause sublethal narcosis in temperate aquatic species. Critical body residues (CBRs) were more reproducible across tests than effective seawater concentrations. These findings support the concept of the CBR approach, that effective tissue residues are comparable across species and geographical ranges despite differences in environmental factors.

Increased feeding and nutrient excretion of adult Antarctic krill, Euphausia superba, exposed to enhanced carbon dioxide (CO2).

Ocean acidification has a wide-ranging potential for impacting the physiology and metabolism of zooplankton. Sufficiently elevated CO(2) concentrations can alter internal acid-base balance, compromising homeostatic regulation and disrupting internal systems ranging from oxygen transport to ion balance. We assessed feeding and nutrient excretion rates in natural populations of the keystone species Euphausia superba (Antarctic krill) by conducting a CO(2) perturbation experiment at ambient and elevated atmospheric CO(2) levels in January 2011 along the West Antarctic Peninsula (WAP). Under elevated CO(2) conditions (∼672 ppm), ingestion rates of krill averaged 78 µg C individual(-1) d(-1) and were 3.5 times higher than krill ingestion rates at ambient, present day CO(2) concentrations. Additionally, rates of ammonium, phosphate, and dissolved organic carbon (DOC) excretion by krill were 1.5, 1.5, and 3.0 times higher, respectively, in the high CO(2) treatment than at ambient CO(2) concentrations. Excretion of urea, however, was ∼17% lower in the high CO(2) treatment, suggesting differences in catabolic processes of krill between treatments. Activities of key metabolic enzymes, malate dehydrogenase (MDH) and lactate dehydrogenase (LDH), were consistently higher in the high CO(2) treatment. The observed shifts in metabolism are consistent with increased physiological costs associated with regulating internal acid-base equilibria. This represents an additional stress that may hamper growth and reproduction, which would negatively impact an already declining krill population along the WAP.

Aqueous uptake and sublethal toxicity of p,p'-DDE in non-feeding larval stages of Antarctic krill (Euphausia superba).

This study evaluated the toxicological sensitivity of non-feeding larval stages of a key Antarctic species (Antarctic krill, Euphausia superba) to p,p'-dichlorodiphenyl dichloroethylene (p,p'-DDE) exposure. The aqueous uptake clearance rate of 84 mL g(-1) preserved weight (p.w.) h(-1) determined for p,p'-DDE in Antarctic krill larvae is comparable to previous findings for small cold water crustaceans and five times slower than the rates reported for an amphipod inhabiting warmer waters. Natural variations in larval physiology appear to influence contaminant uptake and larval krill behavioural responses, strongly highlighting the importance of time of measurement for ecotoxicological testing. Sublethal narcosis (immobility) was observed in larval Antarctic krill from p,p'-DDE body residues of 0.2 mmol/kg p.w., which is in agreement with findings for adult krill and temperate aquatic species. The finding of comparable body residue-based toxicity of p,p'-DDE between polar and temperate species supports the tissue residue approach for environmental risk assessment of polar ecosystems.

Altered developmental timing in early life stages of Antarctic krill (Euphausia superba) exposed to p,p'-DDE.

Persistent organic pollutants (POPs) are persistent, toxic and bioaccumulative anthropogenic organic chemicals, capable of undergoing long range environmental transport to remote areas including the Antarctic. p,p'-dichlorodiphenyl dichloroethylene (p,p'-DDE) has been identified as a dominant POP accumulating in Antarctic krill (Euphausia superba), which is a key Southern Ocean species. This study examined the developmental toxicity of p,p'-DDE via aqueous exposure to Antarctic krill larvae. p,p'-DDE exposure was found to stimulate developmental timing in the first three larval stages of Antarctic krill, while extended monitoring of larvae after a five day exposure period had ended, revealed delayed inhibitory responses during development to the fourth larval stage. Stimulatory responses were observed from the lowest p,p'-DDE body residue tested of 10.1±3.0 µmol/kg (3.2±0.95 mg/kg) preserved wet weight, which is comparable to findings for temperate species and an order of magnitude lower than the exposure level found to cause sublethal behavioural effects in Antarctic krill. The delayed responses included increased mortality, which had doubled in the highest p,p'-DDE treatment (95±8.9% mortality at 20 µg/L p,p'-DDE) compared to the solvent control (44±11% mortality) 2 weeks after end of exposure. Development of surviving metanauplius larvae to calyptopis 1 larvae was delayed by 2 days in p,p'-DDE exposed larvae compared with untreated larvae. Finally, the developmental success of surviving p,p'-DDE exposed larvae was reduced by 50 to 75% compared to the solvent control (100% developmental success). The lowest observed effect concentration for all delayed effects was 1 µg/L, the lowest exposure concentration tested. These findings demonstrate the importance of delayed and indirect effects of toxicant exposure. Further, the findings of this study are important for environmental risk assessment of POPs in the Southern Ocean ecosystem and strongly highlight the significance of developmental endpoints for ecotoxicological testing.

Involvement of contractile elements in control of bioluminescence in Northern krill, Meganyctiphanes norvegica (M. Sars).

Inside the light organs of the bioluminescent (light-producing) crustacean Meganyctiphanes norvegica (krill), numerous capillaries drain haemolymph into the light-producing structure (lantern). We have investigated the arrangement and function of filamentous material found around the opening of the capillaries. These have been suggested to work as sphincters, controlling the haemolymph (i.e. oxygen) supply to the lantern and thereby the production of light. Electron microscopy shows that the filamentous material consists of thick and thin muscle filaments arranged in perpendicular blocks around the opening of each capillary. The actin probe rhodamine phalloidin has revealed that one component is filamentous actin. Clusters of vesicle-dense nerve profiles surround the cells containing filamentous material and antibodies against 5-hydroxytryptamine (5-HT) reveal that 5-HT containing nerves lead to the filamentous area. When exposed to the muscle-relaxing substances papaverine and verapamil, krill respond with luminescence, suggesting that the sphincter structures are functionally involved in the control of light production. Treatment with the muscle-contracting drugs Bay K8544 and thapsigargin gives no light response. Thus, 5-HT stimulates light production in krill; however, a combination of 5-HT and the muscle-relaxing drugs or Bay K8544 potentiates the effect of 5-HT. Thapsigargin quenches the response to 5-HT. Our results corroborate speculations of earlier authors who have suggested that the sphincter structures are of a muscular nature and important in controlling light production in krill. However, other parameters in addition to the oxygen supply to the lantern are involved in controlling bioluminescence in the light organs of M. norvegica.

Persistent organohalogen contaminant burdens in Antarctic krill (Euphausia superba) from the eastern Antarctic sector: a baseline study.

A baseline for persistent organohalogen compound (POC) accumulation in the Antarctic keystone species, Antarctic krill (Euphausia superba) has been established for a 50 degrees longitudinal range of the eastern Antarctic sector. Samples of adult krill, caught from 12 sites distributed between 30 degrees and 80 degrees E (60-70 degrees S), were analysed for >100 organohalogen compounds including chlorinated pesticides, polychlorinated biphenyls (PCBs), polybrominated organic compounds and polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs). Organochlorine pesticides dominated measured krill contaminant burdens with hexachlorobenzene (HCB) as the single most abundant compound quantified. Krill HCB concentrations were comparable to those detected at this trophic level in both the Arctic and temperate northwest Atlantic, lending support for the hypothesis that HCB will approach global equilibrium at a faster rate than other POCs. Para, para'-dichlorodiphenylethene (p,p'-DDE) was detected at notable concentrations. Measurements of DDT and its degradation products provide an important baseline for monitoring the temporal and geographical influence of renewed, DDT usage for malaria-control in affected southern hemisphere countries. In contrast to the Arctic, PCBs did not feature prominently in contaminant burdens of Antarctic krill. The major commercial polybrominated diphenyl ether (PBDE) congeners -99 and -47 were quantified at low background levels with clear concentration spikes observed at around 70 degrees E , in the vicinity of modern, active research stations. The likelihood that local anthropogenic activities are supplementing low PBDE levels, delivered otherwise primarily via long range environmental transport, is discussed. The suspected naturally occurring brominated organic compound, 2,4,6-tribromoanisole (TBA), was a ubiquitous contaminant in all samples whereas the only PCDD/Fs quantifiable were trace levels of octachlorodibenzo-p-dioxin (OCDD) and 1,2,3,4,7,8/1,2,3,4,7,9-hexachlorodibenzofuran (HxCDF). With the aims of; i) Generating a robust and broadly applicable POC auditing platform for the scarcely studied eastern Antarctic sector; ii) Determining the compounds accumulating in Antarctic krill for further toxicity evaluation studies and iii) Establishing a baseline for Antarctic predator exposure to POCs, this study represents one of the most comprehensive reports of POC contamination of the Antarctic food web to date.

Serotonin and nitric oxide interaction in the control of bioluminescence in northern krill, Meganyctiphanes norvegica (M. Sars).

The role of nitric oxide (NO) in the control of bioluminescence (light production) in the crustacean Meganyctiphanes norvegica (krill) was investigated using pharmacological and immunohistochemical methods. All nitrergic drugs tested failed to induce bioluminescence per se but modulated light production stimulated by 5-hydroxytryptamine (5-HT). NO donors [sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP)] injected in live specimens significantly reduced light production stimulated by 5-HT, whereas inhibition of the enzyme NO synthase (NOS) with l-NAME (N(G)-nitro-l-arginine methyl ester) resulted in an enhancement of the 5-HT response. The effects of NO do not seem to be mediated via production of cGMP as injections of a cGMP analogue (8-Bromoguanosine 3',5'-cyclic monophosphate) gave inconclusive effects on the 5-HT-stimulated light response. Inhibition of cGMP production with ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one) did not affect the light response. Moreover, a few individuals showed a considerably higher response to 5-HT in April and June compared with specimens collected in the autumn and winter. Furthermore, both NOS-like and 5-HT-like materials were detected by immunohistochemistry inside the light organs. NOS-like immunoreactivity was primarily observed in structures associated with vessels inside the light organs, whereas 5-HT-like material was abundant in nerve fibres throughout the whole light organ. The results suggest that NO has a modulatory role at several levels in the control of light production in M. norvegica and that NO and 5-HT interact in this regulation.