Evolution of bioluminescence in marine planktonic copepods

Copepods are the dominant taxa in zooplankton communities of the ocean worldwide. Although bioluminescence of certain copepods has been known for more than a 100 years, there is very limited information about the structure and evolutionary history of copepod luciferase genes. Here, we report the cDNA sequences of 11 copepod luciferases isolated from the superfamily Augaptiloidea in the order Calanoida. Highly conserved amino acid residues in two similar repeat sequences were confirmed by the multiple alignment of all known copepod luciferases. Copepod luciferases were classified into two groups of Metridinidae and Heterorhabdidae/Lucicutiidae families based on phylogenetic analyses, with confirmation of the interrelationships within the Calanoida using 18S ribosomal DNA sequences. The large diversity in the specific activity of planktonic homogenates and copepod luciferases that we were able to express in mammalian cultured cells illustrates the importance of bioluminescence as a protective function against predators. We also discuss the relationship between the evolution of copepod bioluminescence and the aspects of their ecological characteristics, such as swimming activity and vertical habitat.     

Sulfate inhibition of molybdate assimilation by planktonic algae and bacteria: some implications for the aquatic nitrogen cycle

Molybdenum is required for both dinitrogen fixation and nitrate assimilation. In oxic waters the primary form of molybdenum is the molybdate anion. Using radioactive [⁹⁹Mol Na₂MoO₄, we have shown that the transport of molybdate by a natural assemblage of freshwater phytoplankton is light-dependent and follows typical saturation kinetics. The molybdate anion is strikingly similar to sulfate and we present data to show that sulfate is a competitive inhibitor of molybdate assimilation by planktonic algae and bacteria. The ability of freshwater phytoplankton to transport molybdate is inhibited at sulfate concentrations as low as 5% of those in seawater and at sulfate: molybdate ratios as low as 50 to 100 times lower than those found in seawater, Similarly, the growth of both a freshwater bacterium and a saltwater diatom was inhibited at sulfate: molybdate ratios lower than those in seawater.The ratio of sulfate to molybdate is 10 to 100 times greater in seawater than in fresh water. This unfavorable sulfate: molybdate ratio may make molybdate less biologically available in the sea. The sulfate: molybdate ratio may explain, in part, the low rates of nitrogen fixation in N-limited salt waters.     

Motion behavior of nauplii and early copepodid stages of marine planktonic copepods

The goal of this study was to quantify periods of activity andvelocities of late naupliar and early copepodid stages of planktoniccopepods occurring regularly on the southeastern continentalshelf of the USA. We obtained quantitative information on eightspecies, including adult females of Oithona plumifera. All studieswere conducted at food concentrations near or above satiationlevels. Activities ranged from 0.85% (adult females of O.plumifera)to 100% of time (nauplii and copepodids of various calanoidspecies). Motion velocities (excluding escape motion) coveredmore than one order of magnitude: from 0.39 mm s^–1 fornauplii of Temora stylifera to 5.24 mm s^–1 for naupliiof Oncaea mediterranea. Ranges of activities of species rangefrom occasional for early juveniles to adult females of O.plumiferato 100% for the same range of T.stylifera, the latter creatinga feeding current from N III onwards, the former not at all.Of notable interest is Centropages velificatus which moves intermittentlyas a late nauplius, continuously as an early copepodid and intermittentlyas an adult. All observed calanoid late nauplii and copepodidsmove in three dimensions, excluding copepodids of the shelfbreak/oceanicParacalanus aculeatus. The results indicate not only significantdifferences in motion behavior between cyclopoids and calanoids,but also between calanoid species. Yet, some calanoid speciesshow little ontogenetic changes at all.     

Feeding, excretion and egg production by individuals and populations of the marine, planktonic copepods, Acartia spp. and Centropages furcatus

Diel variations in vertical distribution, gut pigment content,ammonium excretion and egg production were investigated foradult females of Acartia erythraea and A.pacifica in the verticallymixed Inland Sea of Japan and Centropages furcatus in the stratified,neritic Gulf of Mexico. Gut pigment content and egg productionrate were maximal at night and ammonium excretion was maximalduring the daytime. Neither A.erythraea nor A.pacifica adultfemales showed an apparent diel migration, but the former werehighly concentrated in the surface layer during the afternoon.In contrast, C.furcatus adult females showed a clear diel migration,residing immediately above the bottom during the daytime andbeing concentrated between 10 and 25 m depth during the nighttime.Individual-based data on gut content and excretion and egg productionrates were combined with vertical-distribution data to calculatepopulation values. In the Inland Sea of Japan, the resultantpattern for Acartia spp. reflected the diel variation in physiologicalrates and even distribution of adult females, except for theafternoon, surface aggregation of A.erythraea. In the Gulf ofMexico, the pattern for C.furcatus reflected largely the dielvariation in each rate process and the heterogeneous distributionof adult females in the water column. Elevated nocturnal feedingactivity of these copepods may be due to an endogenous rhythm.The daytime maximum in ammonium excretion and night-time maximumin egg production rate indicated approximate half-day and daytime lags, respectively, after the intake of food until itsconversion into dissolved excreta and released eggs.     

Stable carbon isotope fractionation by marine phytoplankton during photosynthesis

Abstract. In the marine environment, the range of values of carbon isotope fractionation between particulate tissue of phytoplankton and inorganic carbon can be more than 20‰ (− 35‰ < δ¹³C < − 14‰). This review considers the influence of seawater temperature, lipid content of phytoplanktonic cells, kinetic fractionation, and carbon pathway on δ¹³C values observed at sea.
In order to study the contribution of carboxylases (RUBISCO and the β-carboxylases phosphoenolpyruvate carboxylase, phosphoenoplpyruvate carboxykinase and pyruvate carboxylase) to variations of particulate δ¹³C values at sea, we present results obtained simultenously on carboxylase activities and δ¹³C in various environmental conditions. The lowest δ¹³C values are clearly associated with predominance of ribulose-1.5-bisphosphate carboxylase activity, but it was more difficult to explain the high δ¹³C values. Different hypotheses are discussed.     

Bioturbation: impact on the marine nitrogen cycle

Sediments play a key role in the marine nitrogen cycle and can act either as a source or a sink of biologically available (fixed) nitrogen. This cycling is driven by a number of microbial remineralization reactions, many of which occur across the oxic/anoxic interface near the sediment surface. The presence and activity of large burrowing macrofauna (bioturbators) in the sediment can significantly affect these microbial processes by altering the physicochemical properties of the sediment. For example, the building and irrigation of burrows by bioturbators introduces fresh oxygenated water into deeper sediment layers and allows the exchange of solutes between the sediment and water column. Burrows can effectively extend the oxic/anoxic interface into deeper sediment layers, thus providing a unique environment for nitrogen-cycling microbial communities. Recent studies have shown that the abundance and diversity of micro-organisms can be far greater in burrow wall sediment than in the surrounding surface or subsurface sediment; meanwhile, bioturbated sediment supports higher rates of coupled nitrification-denitrification reactions and increased fluxes of ammonium to the water column. In the present paper we discuss the potential for bioturbation to significantly affect marine nitrogen cycling, as well as the molecular techniques used to study microbial nitrogen cycling communities and directions for future study.     

Effect of rhizobial strain and host plant on nitrogen isotopic fractionation in legumes

Lotus pedunculatus L., Medicago sativa L., Macroptilium atropurpureum, Glycine max, and Trifolium repens L. were grown in a N-free medium and inoculated with one of ten Rhizobium strains. Dry matter, N content, and δ¹⁵N values were determined for various plant parts.

Nodules, with the exception of those from lucerne, were enriched in ¹⁵N relative to atmospheric N. Considerable variation was found in δ¹⁵N values of plant herbage (−4.5 to +0.8). The extent of isotopic discrimination was dependent on both the host plant and the infecting rhizobial strain. This further complicates, but does not invalidate, the use of small variations in the natural abundance of ¹⁵N to estimate the contribution of symbiotically fixed N₂ to the N in legume herbage. Some other implications of the observed differences are also discussed.

     

Toxicity of chromium to the marine planktonic copepodAcartia clausi,Giesbrecht

The toxicity of chromium to the marine planktonic copepodAcartia clausi, Giesbrecht was studied. The LC50 48 h values (concentration of chromium lethal to 50% of the test animals) vary with the experimental temperature, the form of chromium compound tested and the annual generation to which theAcartia specimens belong. The elevation of temperature resulted in a considerable increase ofAcartia's sensitivity to chromium. Cr⁶⁺ in the form of Na₂CrO₄ was more toxic toA. clausi than in the form of CrO₃ · Cr³⁺ in the form of Cr(NO₃)₃ 9 H₂O precipitated to the bottom and was not toxic toAcartia.Acartia specimens belonging to the summer generation were more sensitive to chromium than those belonging to the winter or autumn generation. The exposure ofAcartia to sublethal concentrations of chromium resulted in a reduction of its longevity proportional to the chromium concentrations used.Furthermore, whenA. clausi was exposed to sublethal chromium concentrations it showed a decrease of feeding capacity and increase of respiratory rates, which became more pronounced with increasing chromium concentrations.     

Life in transition: balancing inertial and viscous forces by planktonic copepods

Copepods (1-10 mm aquatic crustaceans moving at 1-1000 mm s(-1)) live at Reynolds numbers that vary over 5 orders of magnitude, from 10(-2) to 10(3). Hence, they live at the interface between laminar and turbulent regimes and are subject to the physical constraints imposed by both viscous and inertial realms. At large scales, the inertially driven system enforces the dominance of physically derived fluid motion; plankton, advected by currents, adjust their life histories to the changing oceanic environment. At Kolmogorov scales, a careful interplay of evenly matched forces of biology and physics occurs. Copepods conform or deform the local physical environment for their survival, using morphological and behavioral adaptations to shift the balance in their favor. Examples of these balances and transitions are observed when copepods engage in their various survival tasks of feeding, predator avoidance, mating, and signaling. Quantitative analyses of their behavior give measures of such physical properties of their fluid medium as energy dissipation rates, molecular diffusion rates, eddy size, and eddy packaging. Understanding the micromechanics of small-scale biological-physical-chemical interactions gives insight into factors influencing large-scale dynamics of copepod distribution, patchiness, and encounter probabilities in the sea.     

Fishing impacts on the marine inorganic carbon cycle

1.  Teleost fish excrete precipitated carbonate and make significant contributions to the marine inorganic carbon cycle at regional and global scales. As total carbonate production is linked to fish size and abundance, fishing is predicted to affect carbonate production by modifying fish abundance and size-structure.
2.  We draw on concepts from physiology, metabolic ecology, life history theory, population dynamics and community ecology to develop, validate and apply analytical tools to assess fishing impacts on carbonate production. Outputs suggest that population and community carbonate production fall rapidly at lower rates of fishing than those used as management targets for sustainable yield.
3.  Theoretical predictions are corroborated by estimated trends in carbonate production by a herring population and a coral reef fish community subject to fishing. Our analytical results build on widely applicable relationships between life history parameters and metabolic rates, and can be generalized to most fished ecosystems.
4.   Synthesis and applications . If the maintenance of chemical processes as well as biological process were adopted as a management objective for fisheries then the methods we have developed can be applied to assess the effects of fishing on carbonate production and to advise on acceptable rates of fishing. Maintenance of this ecosystem service would require lower rates of fishing mortality than those recommended to achieve sustainable yield.     

Infection of marine copepods by Metschnikowia sp.

Summary It was observed that yeasts belonging to the genusMetschnikowia parasitize marine copepods both under natural and experimental conditions. The physiological activity ofMetschnikowia sp. was investigated under a variety of conditions and the importance of these organisms in the ecology of the Strait of Georgia has been discussed.     

Differences in nitrogen and carbon stable isotopes between planktonic and benthic microalgae

We compiled published data on the nitrogen and carbon stable isotope ratios of phytoplankton and benthic microalgae from lentic systems and explored the primary factors determining the isotope values among systems. Also, we investigated seasonal changes in nitrogen stable isotope ratios of phytoplankton and benthic microalgae in the strongly acidic lake, Lake Katanuma, which has only two dominant species, Pinnularia acidojaponica as a benthic diatom and Chlamydomonas acidophila, a planktonic green alga. From the published dataset, it may be concluded that δ¹³C of benthic diatoms were more enriched than those of phytoplankton at the same sites, although the nitrogen isotope of phytoplankton and benthic microalgae were similar. This differences in δ¹³C between benthic microalgae and phytoplankton could be explained by the boundary layer effect. On the other hand, nitrogen isotope values of both benthic microalgae and phytoplankton were primarily controlled by the same environmental factor, and boundary layer effects are not the primary factor determining the nitrogen isotope values of microalgae. Also, we showed temporal dynamics in nitrogen isotopes of benthic and planktonic microalgae species in Lake Katanuma, and the trends of nitrogen isotopes are similar between benthic and planktonic microalgae, as concluded from the published dataset.     

Net and gross incorporation of nitrogen by marine copepods fed on N-labelled diatoms: Methodology and trophic studies

The stable isotope of nitrogen (¹⁵N) and an appropriate three-compartment model were used in two 24-h lasting feeding experiments to trace the flow of N through the copepod Acartia discaudata and Calanus helgolandicus fed on ¹⁵N-labelled Skeletonema costatum and Thalassiosira weissflogii, respectively. Details of the labelling technique and principles of the computation of N transport rates are given. At the end of a single 24-h feeding period only about one third of the total amount of N ingested by A. discaudata was incorporated into the copepod's body N; we refer to this rate as net incorporation. Most of the N ingested was lost as ammonium (48% of total N ingested), followed by losses in the form of eggs + fecal pellets (13%) and dissolved organic N (DON, 9%). The sum of net incorporation and the latter losses is defined as gross incorporation. Net incorporation by C. helgolandicus and N losses did not vary over time during a 24 h lasting time-series feeding experiment. On average, 79% of total N ingested was actually incorporated by the copepod whereas mean N losses as ammonium, eggs + fecal pellets represented only 12 and 9%, respectively. After a 24-h feeding period only 2% of N ingested was lost as DON. Inspection of individual DON pathways showed that both A. discaudata and C. helgolandicus highly contributed to total DON production via direct excretion (79 and 64%, respectively). The remaining DON appearing in the DON pool was derived from phytoplankton via direct release and/or indirect release (copepod ‘sloppy feeding’).     

Propagation of planktonic copepods: production and mortality of eggs

Data on fecundity and egg mortality of neritic copepods were collected in various seasons, areas and under various hydrographical conditions. On a seasonal basis variations in fecundity (F) were related to temperature rather than to the abundance of phytoplankton (P). However, a strong correlation between F and P was evident when water column stability varied horisontally or temporally (i.e. at a tidal front or subsequent to a storm). Estimated specific egg-mortalities were variable and occasionally very severe, up to 9.1 d^–1, implying that down to 10^–4% of the eggs survive to hatching. The implications for phenology and distribution of copepods are discussed.     

Sloppy feeding in marine copepods: prey-size-dependent production of dissolved organic carbon

An equation predicting dissolved organic carbon (DOC) productionby sloppy feeding was developed from literature data on apparentgross growth efficiency and copepod-to-prey size ratio basedon the assumption that real gross growth efficiency is independentof relative prey size. The equation suggests that, for copepod-to-preysize ratios <55, DOC production by sloppy feeding can bequantitatively important. It can be calculated as: Q = 0.714– 0.013 x (ESD_copepod/ESD_prey), where Q is the fractionof carbon removed from suspension that is lost as DOC and ESDis equivalent spherical diameter. Predictions by the equationwere corroborated by actual measurements of copepod DOC productionfrom the literature.     

Trimethylamine and trimethylamine N-oxide are supplementary energy sources for a marine heterotrophic bacterium: implications for marine carbon and nitrogen cycling

Bacteria of the marine Roseobacter clade are characterised by their ability to utilise a wide range of organic and inorganic compounds to support growth. Trimethylamine (TMA) and trimethylamine N-oxide (TMAO) are methylated amines (MA) and form part of the dissolved organic nitrogen pool, the second largest source of nitrogen after N₂ gas, in the oceans. We investigated if the marine heterotrophic bacterium, Ruegeria pomeroyi DSS-3, could utilise TMA and TMAO as a supplementary energy source and whether this trait had any beneficial effect on growth. In R. pomeroyi, catabolism of TMA and TMAO resulted in the production of intracellular ATP which in turn helped to enhance growth rate and growth yield as well as enhancing cell survival during prolonged energy starvation. Furthermore, the simultaneous use of two different exogenous energy sources led to a greater enhancement of chemoorganoheterotrophic growth. The use of TMA and TMAO primarily as an energy source resulted in the remineralisation of nitrogen in the form of ammonium, which could cross feed into another bacterium. This study provides greater insight into the microbial metabolism of MAs in the marine environment and how it may affect both nutrient flow within marine surface waters and the flux of these climatically important compounds into the atmosphere.