Mass spectroscopy reveals compositional differences in copepodamides from limnic and marine copepods.

Marine copepods, the most abundant animals in the global ocean, imprint their surrounding waters with chemical cues, called copepodamides. Copepodamides induce defensive traits such as toxin production, bioluminescence, and colony size plasticity in a variety of marine phytoplankton. The role of copepodamides in freshwater ecosystems is, however, unknown. Here we report the consistent presence of copepodamides in copepods from six Swedish freshwater lakes. Copepodamide concentrations in freshwater copepods are similar to those of marine copepods, around 0.1 ppt of dry mass in millimetre sized individuals. The composition substantially overlaps with marine copepodamides but is also distinctly different. Marine copepods commonly contain both subgroups of copepodamides, the copepodamides (CA) and the dihydro-copepodamides (dhCA), whereas freshwater copepods are dominated by the dhCAs. Taxonomic groups had consistent copepodamide profiles across sampling sites and timepoints, supporting the presence of species-specific copepodamide signatures. We describe 10 new copepodamide structures, four of which were found exclusively in freshwater copepods. The presence of copepodamides in limnic copepods also warrants studies into their potential function as predator alarm cues in freshwater systems.

Rapid measurement of total lipids in zooplankton using the sulfo-phospho-vanillin reaction.

Zooplankton provide a vital source of nutrition to a variety of fish and marine predators. Measuring the total lipid content of zooplankton provides important information about diet quality available to predators, revealing details about trophic dynamics and ecosystem status. We analyze the performance of a microplate assay, utilizing the sulfo-phospho-vanillin (SPV) reaction, to quantify the total lipid content of various large crustacean zooplankton in a rapid and high throughput manner. Pilot experiments were performed by measuring the total lipid content of purchased freeze-dried zooplankton (Calanus finmarchicus and Euphausia superba) by both SPV and gravimetric analysis (low throughput and requires large sample size). The results of the SPV assay were not statistically different from gravimetric analysis for either species (p > 0.05). Further, an inter-laboratory comparison study was performed to measure the total lipid content (% of wet mass) of field-collected Arctic and North Pacific zooplankton (copepods (n = 19) and euphausiids (n = 29)) of various species utilizing multiple analysis methods. Results from thin layer chromatography with flame ionization detection (TLC-FID) demonstrated that lipid classes in zooplankton samples varied in composition of steryl/wax esters (3-95%), triacylglycerols (1-52%), free-fatty acids (0.4-25%), sterols (0-4%) and polar lipids (1-42%). Despite this variation in lipid class composition among samples, the results of the SPV assay agreed well with gravimetric analysis. The mean absolute and relative differences between SPV and gravimetric analysis for all zooplankton lipids in this study were 1.0% and 11.6%, respectively. The SPV assay is rapid (<2 hours), high throughput (25 samples processed in parallel), low cost (supplies <$ 0.67 per sample), precise (inter assay CV = 6.9%, intra assay CV = 6.0%), sensitive (limit of detection < 1.7 micrograms of lipid per analysis), and accurate when calibrated with appropriate standards.

Marine Oil from C. finmarchicus Enhances Glucose Homeostasis and Liver Insulin Resistance in Obese Prediabetic Individuals.

The intermediate state between normal glucose tolerance and overt type 2 diabetes mellitus is associated with micro- and macrovascular diseases, requiring safe and cost-effective treatment measures interventions. A novel source of LC n-3 FAs is Calanus finmarchicus Oil, which showed promising effects on glucose homeostasis in preclinical studies due to anti-obesity effects and/or anti-inflammatory properties. In total, 43 obese patients (BMI: 31.7 ± 5.2 kg/m2) were allocated in the following two groups: (1) Calanus oil group (2 g CO/day) and (2) placebo group (2 g paraffin oil/day). Markers of glucose metabolism, body composition and energy intake were measured at the beginning (t0), after 12 weeks (t12) and 16 weeks (t16). Overall, parameters reflecting abnormal glucose homeostasis and insulin resistance in the liver, including fasting insulin (-2.9 mU/L ± 4.10, p < 0.05), HOMA-IR (-0.9 ± 1.28, p < 0.05) and hepatic insulin resistance index (-1.06 ± 1.72 × 106, p < 0.05) significantly enhanced after a 12-week CO-intervention, while no differences were observed in HbA1c, AUC0-2h Glucose, AUC0-2h Insulin, 2 h plasma glucose and muscle insulin sensitivity index. Our results indicate that Calanus oil causes beneficial effects on glucose metabolism and insulin resistance in obese patients, with clinical relevance to be verified in further studies. In addition, the possible active compounds and their mechanisms of action should be elucidated.

Polar and neutral lipid composition of the copepod Lernaeocera lusci and its host Merluccius merluccius in relationship with the parasite intensity.

Parasitic copepod Lernaeocera lusci is a common mesoparasite of the hake Merluccius merluccius. Although widely distributed throughout the Mediterranean, little is known about this pathogen. The current study was designed to assess the impact of different L. lusci infection loads on lipid classes and their fatty acid (FA) composition in both parasite and the host organs (gills, liver, and muscle). Results showed a significant decrease in total lipid, neutral lipid (NL), and polar lipid (PL) contents in all analyzed host's organs in relationship with parasite intensity. Gills appeared to be the most impacted organ under the lowest parasite intensity (loss of 50% of NL and PL amounts). At the highest parasitic infection, a loss of about 80% of lipid moieties was recorded in all analyzed organs. Simultaneously, no significant differences were found for the parasite reflecting its ability to sustain an appropriate lipid amount required for its survival and development. Significant changes in the FA composition were recorded in both host and parasite. Particularly, we have noticed that for L. lusci, the intraspecific competition has resulted in an increased level of some essential FA such as C22:6n-3 (docosahexaenoic acid, DHA), C20:5n-3 (eicosapentaenoic acid, EPA), and C20:4n-6 (arachidonic acid, ARA). This probably reflects that in addition to a direct host FA diversion, L. Lusci can modulate its FA composition by increasing the activity of desaturation. Within the host, liver PL appeared to be the less impacted fraction which may mirror an adaptive strategy adopted by the host to preserve the structural and functional integrity of this vital organ.

Microbial dynamics of elevated carbon flux in the open ocean's abyss.

In the open ocean, elevated carbon flux (ECF) events increase the delivery of particulate carbon from surface waters to the seafloor by severalfold compared to other times of year. Since microbes play central roles in primary production and sinking particle formation, they contribute greatly to carbon export to the deep sea. Few studies, however, have quantitatively linked ECF events with the specific microbial assemblages that drive them. Here, we identify key microbial taxa and functional traits on deep-sea sinking particles that correlate positively with ECF events. Microbes enriched on sinking particles in summer ECF events included symbiotic and free-living diazotrophic cyanobacteria, rhizosolenid diatoms, phototrophic and heterotrophic protists, and photoheterotrophic and copiotrophic bacteria. Particle-attached bacteria reaching the abyss during summer ECF events encoded metabolic pathways reflecting their surface water origins, including oxygenic and aerobic anoxygenic photosynthesis, nitrogen fixation, and proteorhodopsin-based photoheterotrophy. The abundances of some deep-sea bacteria also correlated positively with summer ECF events, suggesting rapid bathypelagic responses to elevated organic matter inputs. Biota enriched on sinking particles during a spring ECF event were distinct from those found in summer, and included rhizaria, copepods, fungi, and different bacterial taxa. At other times over our 3-y study, mid- and deep-water particle colonization, predation, degradation, and repackaging (by deep-sea bacteria, protists, and animals) appeared to shape the biotic composition of particles reaching the abyss. Our analyses reveal key microbial players and biological processes involved in particle formation, rapid export, and consumption, that may influence the ocean's biological pump and help sustain deep-sea ecosystems.

Intake of Calanus finmarchicus oil for 12 weeks improves omega-3 index in healthy older subjects engaging in an exercise programme.

The n-3 PUFA, EPA and DHA, play an important role in human health. As the intake of EPA and DHA from the diet is often inadequate, supplementation of those fatty acids is recommended. A novel source of n-3 PUFA is Calanus finmarchicus oil (CO) which contains fatty acids mainly bound in wax esters. To date, no data are available on the effects of long-term intake of this marine oil on n-3 PUFA blood levels. Therefore, the aim of this study was to evaluate the effect of CO on the n-3 PUFA blood levels using the omega-3 index (O3I). The data originate from a larger randomised controlled trial. For this analysis, samples from seventy-two participants (59·2 (sd 6·2) years, BMI 27·7 (sd 5·28) kg/m2) were analysed. Of those, thirty-six performed 2×/week exercise and received 2 g of CO, which provided 124 mg stearidonic acid (SDA), 109 mg EPA and 87 mg DHA daily (EXCO group), while the other group performed exercise only (EX group) and served as a control for this analysis. The O3I increased from 6·07 (sd 1·29) % at baseline to 7·37 (sd 1·10) % after 12 weeks within the EXCO group (P < 0·001), while there were no significant changes in the EX group (6·01 (sd 1·26)-6·15 (sd 1·32) %, P = 0·238). These data provide first evidence that wax ester-bound n-3 PUFA from CO can significantly increase the O3I despite relatively low EPA + DHA amounts. Further, the effects of exercise could be excluded.

Effects of Exercise Combined with a Healthy Diet or Calanus finmarchicus Oil Supplementation on Body Composition and Metabolic Markers-A Pilot Study.

Aging is accompanied by a progressive decline in muscle mass and an increase in fat mass, which are detrimental changes associated with the development of health conditions such as type-2 diabetes mellitus or chronic low-grade inflammation. Although both exercise as well as nutritional interventions are known to be beneficial in counteracting those age-related changes, data to which extent untrained elderly people may benefit is still sparse. Therefore, a randomized, controlled, 12-week interventional trial was conducted in which 134 healthy untrained participants (96 women and 38 men, age 59.4 ± 5.6 years, body mass index (BMI) 28.4 ± 5.8 kg/m2) were allocated to one of four study groups: (1) control group with no intervention (CON); (2) 2×/week aerobic and resistance training only (EX); (3) exercise routine combined with dietary counseling in accordance with the guidelines of the German Nutrition Society (EXDC); (4) exercise routine combined with intake of 2 g/day oil from Calanus finmarchicus (EXCO). Body composition (bioelectrical impedance analysis), as well as markers of glucose metabolism and blood lipids, were analyzed at the beginning and the end of the study. The highest decreases in body fat were observed within the EXCO group (-1.70 ± 2.45 kg, p < 0.001), and the EXDC (-1.41 ± 2.13 kg, p = 0.008) group. Markers of glucose metabolism and blood lipids remained unchanged in all groups. Taken together results of this pilot study suggest that a combination of moderate exercise and intake of oil from Calanus finmarchicus or a healthy diet may promote fat loss in elderly untrained overweight participants.

The potential of fatty acid isotopes to trace trophic transfer in aquatic food-webs.

Compound-specific isotope analyses (CSIA) of fatty acids (FA) constitute a promising tool for tracing energy flows in food-webs. However, past applications of FA-specific carbon isotope analyses have been restricted to a relatively coarse food-source separation and mainly quantified dietary contributions from different habitats. Our aim was to evaluate the potential of FA-CSIA to provide high-resolution data on within-system energy flows using algae and zooplankton as model organisms. First, we investigated the power of FA-CSIA to distinguish among four different algae groups, namely cyanobacteria, chlorophytes, haptophytes and diatoms. We found substantial within-group variation but also demonstrated that δ13C of several FA (e.g. 18:3ω3 or 18:4ω3) differed among taxa, resulting in group-specific isotopic fingerprints. Second, we assessed changes in FA isotope ratios with trophic transfer. Isotope fractionation was highly variable in daphnids and rotifers exposed to different food sources. Only δ13C of nutritionally valuable poly-unsaturated FA remained relatively constant, highlighting their potential as dietary tracers. The variability in fractionation was partly driven by the identity of food sources. Such systematic effects likely reflect the impact of dietary quality on consumers' metabolism and suggest that FA isotopes could be useful nutritional indicators in the field. Overall, our results reveal that the variability of FA isotope ratios provides a substantial challenge, but that FA-CSIA nevertheless have several promising applications in food-web ecology. This article is part of the theme issue 'The next horizons for lipids as 'trophic biomarkers': evidence and significance of consumer modification of dietary fatty acids'.

Effect of Calanus Oil Supplementation and 16 Week Exercise Program on Selected Fitness Parameters in Older Women.

We investigated changes in functional fitness after an exercise program in combination with Calanus oil supplementation, a novel source of bioactive lipids rich in wax esters with omega-3 polyunsaturated fatty acid (n-3 PUFA). Fifty-five healthy sedentary women aged 65-80 (mean age 70.9 ± 3.9 years, BMI 27.24 ± 3.9 kg m-2, VO2peak 19.46 ± 3.7 ml kg-1 min-1) were enrolled in the study. The participants were divided into two groups: exercise training plus Calanus Oil supplementation (n = 28) or exercise plus placebo (sunflower oil) supplementation (n = 27). The exercise intervention program was completed by 53 participants and contained functional circuit training (twice a week, 45 min plus 15 min of stretching and balance training) and Nordic walking (once a week, 60 min) for 16 weeks. Senior fitness test, exercise stress test on bicycle ergometer, hand-grip, and body composition were evaluated before and after the program. Our results show that functional fitness and body composition improved following the interventional exercise program, but for most of the parameters there was no synergic effect of supplementing n-3 PUFA-rich Calanus oil. In comparison to the placebo group, the group with Calanus supplementation experienced significantly higher improvement of functional strength of lower body which was evaluated by the chair stand test. Supplementation with Calanus may have a synergic effect with exercise on functional strength of the lower body in the elderly.

Calanus oil in the treatment of obesity-related low-grade inflammation, insulin resistance, and atherosclerosis.

Calanus oil (COil) is a natural product extracted from marine zooplankton Calanus finmarchicus found in the North Atlantic Ocean. This oil is rich in wax esters of polyunsaturated fatty acids (PUFAs) and has been projected as the best alternative to fish oil because its production cannot keep pace with the demands from the growing markets. The COil is the only commercially available marine source of wax esters, whereas classic ω-3 PUFAs comes from triglycerides, ethyl esters, and phospholipids. It has, in recent decades, been seen that there is an unprecedented rise in the use of PUFA-rich oil in the aquaculture industry. A simultaneous rise in the demand of PUFAs is also observed in the health care industry, where PUFAs are suggested preventing various disorders related to lifestyles such as obesity, diabetes mellitus, chronic low-grade inflammation, atherosclerosis, and brain and cardiovascular disorders (CVDs). In this review, we will explore the metabolic aspects related to the use of COil as an antioxidant, anticholesterinemic, and anti-inflammatory dietary source and its impact on the prevention and therapy of obesity-related metabolic disorders.

A scavenger receptor B (CD36)-like protein is a potential mediator of intestinal heme absorption in the hematophagous ectoparasite Lepeophtheirus salmonis.

Intestinal absorption of heme has remained enigmatic for years, even though heme provides the most bioavailable form of iron. The salmon louse, Lepeophtheirus salmonis, is a heme auxotrophic ectoparasite feeding on large quantities of blood from its host, the salmon. Here we show that a scavenging CD36-like receptor is a potential mediator of heme absorption in the intestine of the salmon louse. The receptor was characterized by a heme binding assay using recombinantly expressed protein, in situ hybridization and immunohistochemistry, as well as functional knockdown studies in the louse. A computational structural model of the receptor predicted a binding pocket for heme, as also supported by in silico docking. The mRNA and protein were expressed exclusively in the intestine of the louse. Further, knocking down the transcript resulted in lower heme levels in the adult female louse, production of shorter egg strings, and an overall lower hatching success of the eggs. Finally, starving the lice caused the transcript expression of the receptor to decrease. To our knowledge, this is the first time a CD36-like protein has been suggested to be an intestinal heme receptor.

Seasonal and spatial fatty acid profiling of the calanoid copepods Temora longicornis and Acartia clausi linked to environmental stressors in the North Sea.

The Belgian part of the North Sea (BPNS) is subjected to multiple environmental stressors. The impact of these stressors includes the modulation of fatty acid (FA) composition of the zooplankton. This study recorded temporal and spatial patterns of the FA profiles of two dominant calanoid copepods within the BPNS: Temora longicornis (Müller, 1785) and Acartia clausi (Giesbrecht, 1889). By means of distance-based linear modelling and by applying multi model inference to generalized additive models, environmental stressors were linked to patterns of the FA profiles of these species. The FA profiles of A. clausi and T. longicornis showed distinct intraspecific, spatial and temporal differences within the BPNS. Temperature and algal food quality (marked by the ratio of silicate concentration to dissolved inorganic nitrogen concentration, SiO4/DIN) were the most important drivers of seasonal fluctuations in the DHA/EPA ratio of both species. DHA/EPA ratio can be used as marker for stress in copepods in the BPNS in order to have a quick indication of food quality changes at the basis of the food web.

Embryos of an Antarctic zooplankton require anoxia for dormancy, are permeable to lipophilic chemicals, and reside in sediments containing PCBs.

Zooplankton in Antarctic maritime lakes face challenges imposed by anthropogenic chemicals. Studies on temperate species suggest that lipophilic chemicals will accumulate in dormant embryos of Antarctic zooplankton and decrease hatching success, thereby threatening centuries of accumulated genetic diversity that would increase population resilience in the face of climate change. We evaluated the potential for lakes to act as sinks for legacy pollutants in the maritime Antarctic by testing sediments for polychlorinated biphenyls (PCBs) previously identified in soil, flora and fauna of lake catchments. Direct tests of embryo permeability to chemicals are confounded by potential adhesion of chemicals to the embryo surface and limited biomass available. Therefore, in order to assess the potential for lipophilic chemicals to penetrate and passively accumulate in dormant embryos of Antarctic lacustrine zooplankton, we evaluated the effect of anoxia on post-diapause development in the calanoid copepod, Boeckella poppei, and then used chemical anoxia induced by rotenone as a reporter for permeability of these embryos to moderately lipophilic chemicals. The data presented demonstrate that embryos of B. poppei from Antarctic lake sediments will passively accumulate moderately lipophilic chemicals while lying dormant in anoxic sediments. Implications for legacy POPs in sediments of Antarctic maritime lakes are discussed.

210Po concentration in selected calanoid copepods in the northern Arabian Gulf.

Copepods are the most abundant metazoans, forming a vital food chain link between the primary producers the phytoplankton and fish. This study presents baseline information on the concentration of 210Po among calanoid copepods isolated from the Kuwait marine area. The concentration of 210Po in six species of copepod, including Subeucalanus flemingeri, Parvocalanus crassirostis, Acartia pacifica, Calanopia elliptica, Acrocalanus gibber, and Euterpina acutifrons were 151.3-158.8 Bq kg-1 wwt, 121.1-129.5 Bq kg-1 wwt, 51.23-54.91 Bq kg-1 wwt, 38.88-40.09 Bq kg-1 wwt, 38.07-38.29 Bq kg-1 wwt, and 33.46-36.50 Bq kg-1 wwt, respectively. The 210Po concentration in seawater shows a seasonal variation, with a higher concentration range of 0.58-0.70 mBq L-1 during summer and autumn, while a lower concentration is found (0.30-0.38 mBq L-1) during winter and spring. The concentration factor among the copepods varies between 8 ∗ 104 and 5 ∗ 105 that is an order of magnitude higher than the diatoms and dinoflagellates.

Characterisation of proteins in excretory/secretory products collected from salmon lice, Lepeophtheirus salmonis.

BACKGROUND: The salmon louse, Lepeophtheirus salmonis, is an ectoparasitic copepod which feeds on the mucus, skin and blood of salmonid fish species. The parasite can persist on the surface of the fish without any effective control being exerted by the host immune system. Other ectoparasitic invertebrates produce compounds in their saliva, excretions and/or secretions which modulate the host immune responses allowing them to remain on or in the host during development. Similarly, compounds are produced in secretions of L. salmonis which are thought to be responsible for immunomodulation of the host responses as well as other aspects of crucial host-parasite interactions. METHODS: In this study we have identified and characterised the proteins in the excretory/secretory (E/S) products of L. salmonis using LC-ESI-MS/MS. RESULTS: In total 187 individual proteins were identified in the E/S collected from adult lice and pre-adult sea lice. Fifty-three proteins, including 13 serine-type endopeptidases, 1 peroxidase and 5 vitellogenin-like proteins were common to both adult and pre-adult E/S products. One hundred and seven proteins were identified in the adult E/S but not in the pre-adult E/S and these included serine and cysteine-type endopeptidases, vitellogenins, sphingomyelinase and calreticulin. A total of 27 proteins were identified in pre-adult E/S products but not in adult E/S. CONCLUSIONS: The assigned functions of these E/S products and the potential roles they play in host-parasite interaction is discussed.

Managing wastes as green resources: cigarette butt-synthesized pesticides are highly toxic to malaria vectors with little impact on predatory copepods.

The development of novel mosquito control tools is a key prerequisite to build effective and reliable Integrated Vector Management strategies. Here, we proposed a novel method using cigarette butts for the synthesis of Ag nanostructures toxic to young instars of the malaria vector Anopheles stephensi, chloroquine (CQ)-resistant malaria parasites Plasmodium falciparum and microbial pathogens. The non-target impact of these nanomaterials in the aquatic environment was evaluated testing them at sub-lethal doses on the predatory copepod Mesocyclops aspericornis. Cigarette butt-synthesized Ag nanostructures were characterized by UV-vis and FTIR spectroscopy, as well as by EDX, SEM and XRD analyses. Low doses of cigarette butt extracts (with and without tobacco) showed larvicidal and pupicidal toxicity on An. stephensi. The LC50 of cigarette butt-synthesized Ag nanostructures ranged from 4.505 ppm (I instar larvae) to 8.070 ppm (pupae) using smoked cigarette butts with tobacco, and from 3.571 (I instar larvae) to 6.143 ppm (pupae) using unsmoked cigarette butts without tobacco. Smoke toxicity experiments conducted against adults showed that unsmoked cigarette butts-based coils led to mortality comparable to permethrin-based positive control (84.2 and 91.2%, respectively). A single treatment with cigarette butts extracts and Ag nanostructures significantly reduced egg hatchability of An. stephensi. Furthermore, the antiplasmodial activity of cigarette butt extracts (with and without tobacco) and synthesized Ag nanostructures was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of P. falciparum. The lowest IC50 values were achieved by cigarette butt extracts without tobacco, they were 54.63 µg/ml (CQ-s) and 63.26 µg/ml (CQ-r); while Ag nanostructure IC50 values were 72.13 µg/ml (CQ-s) and 77.33 µg/ml (CQ-r). In MIC assays, low doses of the Ag nanostructures inhibited the growth of Bacillus subtilis, Klebsiella pneumoniae and Salmonella typhi. Finally, the predation efficiency of copepod M. aspericornis towards larvae of An. stephensi did not decrease in a nanoparticle-contaminated environment, if compared to control predation assays. Overall, the present research would suggest that an abundant hazardous waste, such as cigarette butts, can be turned to an important resource for nanosynthesis of highly effective antiplasmodials and insecticides.

Engineering Intracellularly Retained Gaussia Luciferase Reporters for Improved Biosensing and Molecular Imaging Applications.

Gaussia luciferase (GLUC) is a bioluminescent reporter protein of increasing importance. As a secretory protein, it has increased sensitivity in vitro and in vivo (∼20 000-fold, and ∼1000-fold, respectively) over its competitor, secreted alkaline phosphatase. Unfortunately, this same advantageous secretory nature of GLUC limits its usefulness for many other possible intracellular applications, e.g., imaging signaling pathways in intact cells, in vivo imaging, and in developing molecular imaging biosensors to study protein-protein interactions and protein folding. Hence, to widen the research applications of GLUC, we developed engineered variants that increase its intracellular retention both by modifying the N-terminal secretory signal peptide and by tagging additional sequences to its C-terminal region. We found that when GLUC was expressed in mammalian cells, its N-terminal secretory signal peptide comprising amino acids 1-16 was essential for GLUC folding and functional activity in addition to its inherent secretory property. Modification of the C-terminus of GLUC by tagging a four amino acid (KDEL) endoplasmic reticulum targeting peptide in multiple repeats significantly improved its intracellular retention, with little impact on its folding and enzymatic activity. We used stable cells expressing this engineered GLUC with KDEL repeats to monitor chemically induced endoplasmic reticulum stress on cells. Additionally, we engineered an apoptotic sensor using modified variants of GLUC containing a four amino acid caspase substrate peptide (DEVD) between the GLUC protein and the KDEL repeats. Its use in cell culture resulted in increased GLUC secretion in the growth medium when cells were treated with the chemotherapeutic drugs doxorubicin, paclitaxel, and carboplatin. We thus successfully engineered a new variant GLUC protein that is retained inside cells rather than secreted extracellularly. We validated this novel reporter by incorporating it in biosensors for detection of cellular endoplasmic reticulum stress and caspase activation. This new molecularly engineered enzymatic reporter has the potential for widespread applications in biological research.

First investigation of trace metal distribution in surface seawater and copepods of the south coast of Sfax (Tunisia).

The increased metal loading from anthropogenic sources has affected aquatic ecosystems and has cascaded through food webs worldwide. Therefore, the evaluation of ecological impacts of anthropogenic metal has become increasingly important. In this paper, we monitored the concentration of six trace metals (Cd, Cu, Fe, Ni, Pb, and Zn) in the three copepod groups (cyclopoida, calanoida, and harpacticoida) and in seawater samples collected from the south coast of Sfax (Tunisia). Results showed that the concentration of Fe and Zn for all copepod groups was higher than that for other metals and that of Cd was the lowest in all groups. The mean increase in bioconcentration factor of metals in copepods ranged from 0.05 to 18.93 and followed the sequence Zn (18.93) > Fe (14.34) > Pb (6.41) > Cd (1.53) > Cu (0.10) > Ni (0.05). The copepods in the south coast of Sfax were found to have a great capacity to accumulate trace metals and act as contamination indicators. Comparative studies with those from the Luza zone indicate considerable bioaccumulation of trace metals (Pb and Ni) in all copepod groups namely in cyclopoida.

Bioaccumulation of methylmercury in a marine copepod.

Methylmercury (MeHg) is known to biomagnify in marine food chains, resulting in higher concentrations in upper trophic level animals than their prey. To better understand how marine copepods, an important intermediate between phytoplankton and forage fish at the bottom of the food chain, assimilate and release MeHg, the authors performed a series of laboratory experiments using the gamma-emitting radiotracer 203 Hg2+ and Me203 Hg with the calanoid copepod Acartia tonsa. Assimilation efficiencies of Hg2+ and MeHg ranged from 25% to 31% and 58% to 79%, respectively, depending on algal diets. Assimilation efficiencies were positively related to the fraction of Hg in the cytoplasm of the algal cells that comprised their diet. Efflux rates of Hg2+ (0.29/d) and MeHg (0.21/d) following aqueous uptake were similar, but efflux rates following dietary uptake were significantly lower for MeHg (0.11-0.22/d) than Hg2+ (0.47-0.66/d). The calculated trophic transfer factors in copepods were >1 for MeHg and consistently low (≤0.2) for Hg2+ . The authors used the parameters measured to quantitatively model the relative importance of MeHg sources (water or diet) for copepods and to predict the overall MeHg concentrations in copepods in different marine environments. In general, MeHg uptake from the diet accounted for most of the body burden in copepods (>50%). For an algal diet with a MeHg dry weight bioconcentration factor ≥106 , >90% of a copepod's MeHg body burden can be shown to derive from the diet. The model-predicted MeHg concentrations in the copepods were comparable to independent measurements for copepods in coastal and open-ocean regions, implying that the measured parameters and model are applicable to natural waters. Environ Toxicol Chem 2017;36:1287-1293. © 2016 SETAC.

Wax Ester Rich Oil From The Marine Crustacean, Calanus finmarchicus, is a Bioavailable Source of EPA and DHA for Human Consumption.

Oil from the marine copepod, Calanus finmarchicus, which contains >86 % of fatty acids present as wax esters, is a novel source of n-3 fatty acids for human consumption. In a randomized, two-period crossover study, 18 healthy adults consumed 8 capsules providing 4 g of Calanus(®) Oil supplying a total of 260 mg EPA and 156 mg DHA primarily as wax esters, or 1 capsule of Lovaza(®) providing 465 mg EPA and 375 mg DHA as ethyl esters, each with an EPA- and DHA-free breakfast. Plasma EPA and DHA were measured over a 72 h period (t = 1, 2, 4, 6, 8, 10, 12, 24, 48, and 72 h). The positive incremental area under the curve over the 72 h test period (iAUC0-72 h) for both EPA and DHA was significantly different from zero (p < 0.0001) in both test conditions, with similar findings for the iAUC0-24 h and iAUC0-48 h, indicating the fatty acids were absorbed. There was no difference in the plasma iAUC0-72 h for EPA + DHA, or DHA individually, in response to Calanus Oil vs the ethyl ester condition; however, the iAUC0-48 h and iAUC0-72 h for plasma EPA in response to Calanus Oil were both significantly increased relative to the ethyl ester condition (iAUC0-48 h: 381 ± 31 vs 259 ± 39 µg*h/mL, p = 0.026; iAUC0-72 h: 514 ± 47 vs 313 ± 49 µg*h/mL, p = 0.009). These data demonstrate a novel wax ester rich marine oil is a suitable alternative source of EPA and DHA for human consumption.