Marine Planktonic Copepods Research Articles

On the morphology of urosome appendages and their potential function of marine planktonic copepods

Abstract The goal of this paper is to draw the attention of biological oceanographers to the urosome of zooplankton. The methodology to determine the utilization of urosomes and their sensors for predator recognition is in its infancy. We here describe the potential utilization of setae and their activity of mostly juvenile copepods for the purpose of perceiving predators. Our morphological observations let us assume that calanoid nauplii have few means to perceive predators approaching from the rear and most likely from frontal, as their appendages are moving most of the time. This is being compared to post-nauplii, i.e. copepodid stages, and adults who possess a 2D array of setae on their urosomes, potentially able to perceive predators approaching from the rear. Detailed small-scale observations should reveal the sensor potential of urosomes of nauplii and older stages of planktonic copepods.

Latitudinal diversity of planktonic copepods in the Eastern Pacific: overcoming sampling biases and predicting patterns

Gradients of latitudinal diversity are one of the biogeographic features calling the most attention in ecology and macroecology. However, in pelagic communities of the marine environment, geographic trends and patterns are poorly known. We evaluated the latitudinal variation in species richness of marine planktonic copepods in the Eastern Pacific using spatial statistical models and approaches that mitigate and account for biases in occurrence data. A Boosted Regression Tree (BRT) and regression-Kriging based models allowed us to estimate and predict alpha diversity in poorly sampled regions, whereas beta diversity patterns were assessed using generalized dissimilarity analysis (GDM). Species richness showed a bimodal pattern, with a maximum of 291 species in the Northern Hemisphere and Tropical Eastern Pacific Ocean. Particulate Organic Carbon, salinity (max), spatial autocovariate, range of salinity and temperature, and Mixed Layer Depth, explained 85.2% of the latitudinal variability of copepods. Beta diversity was structured into four macrozones associated with the main water masses of the North and South Pacific.Our analytical approaches can overcome the limitations of data gaps, predicting greater diversity in subtropical and coastal areas, while providing insights into key drivers modulating spatial diversity patterns.

Effects of nickel oxide nanoparticles on survival, reproduction, and oxidative stress biomarkers in the marine calanoid copepod Centropages ponticus under short-term exposure.

Excessive use of nickel oxide nanoparticles (NiO NPs) in various industrial and commercial products can lead to various negative effects in human and environmental health due to their possible discharge into the environment. Nerveless, information about their ecotoxicological effects on marine organisms are lacking. Copepods are good ecotoxicological models because of their high sensitivity to environmental stress and their key role in the marine food webs. In this study, 48h acute tests were conducted on the marine planktonic copepod Centropages ponticus to assess lethal and sublethal toxicities of NiO NPs. The results revealed LC50 (48h) of 4mg/L for adult females. Aggregation and settling of NiO NPs were observed at concentrations ≥ 2mg/L. Exposure to sublethal concentrations (≥ 0.02mg/L for 48h) had significant negative effects on reproductive success in C. ponticus. Egg production after 24h and 48h decreased by 32% and 46%, respectively at 0.02mg/L and 70% and 82%, respectively, at 2mg/L. Hatching success was reduced by 70% and 79% at 2mg/L for eggs produced after 24h and 48h respectively. Antioxidant enzymatic activity increased significantly with NiO NP concentration and time, indicating that NiO NPs can cause oxidative stress in C. ponticus even under short-term exposure, while significant inhibition of acetylcholinesterase activity at 2mg/L after 48h suggests neurotoxic effects of NiO NPs.

Warmer temperature increases mercury toxicity in a marine copepod.

Marine biota have been co-challenged with ocean warming and mercury (Hg) pollution over many generations because of human activities; however, the molecular mechanisms to explain their combined effects are not well understood. In this study, a marine planktonic copepod Pseudodiaptomus annandalei was acutely exposed to different temperature (22 and 25°C) and Hg (0 and 118μg/L) treatments in a 24-h cross-factored experiment. Hg accumulation and its subcellular fractions were determined in the copepods after exposure. The expression of the genes of superoxide dismutase (SOD), glutathione peroxidase (GPx), metallothionein1 (mt1), heat shock protein 70 (hsp70), hsp90, hexokinase (hk), and pyruvate kinase (pk) was also analyzed. Both the Hg treatment alone and the combined exposure of warmer temperature plus Hg pollution remarkably facilitated Hg bioaccumulation in the exposed copepods. Compared with the Hg treatment alone, the combined exposure increased total Hg accumulation and also the amount of Hg stored in the metal-sensitive fractions (MSF), suggesting elevated Hg toxicity in P. annandalei under a warmer environment, given that the MSF is directly related to metal toxicity. The warmer temperature significantly up-regulated the mRNA levels of mt1, hsp70, hsp90, and hk, indicating the copepods suffered from thermal stress. With exposure to Hg, the mRNA level of SOD increased strikingly but the transcript levels of hsp90, hk, and pk decreased significantly, indicating that Hg induced toxic events (e.g., oxidative damage and energy depletion). Particularly, in contrast to the Hg treatment alone, the combined exposure significantly down-regulated the mRNA levels of SOD and GPx but up-regulated the mRNA levels of mt1, hsp70, hsp90, hk, and pk. Collectively, the results of this study indicate that ocean warming will potentially boost Hg toxicity in the marine copepod P. annandalei, which is information that will increase the accuracy of the projections of marine ecosystem responses to the joint effects of climate change stressors and metal pollution on the future ocean.

Effects of multigenerational rearing, ontogeny and predation threat on copepod feeding rhythms

Marine planktonic copepods frequently exhibit diel feeding cycles coupled with vertical migrations. However, copepod feeding rhythms can be influenced by factors others than different food availability between layers. In this study, we determined the changes in the diel feeding behaviour of two marine copepod species (Centropages typicus and Paracartia grani) through multigenerational laboratory rearing, ontogeny, and upon the exposure to predator chemical cues. The wild females of both C. typicus and P. grani showed marked feeding rhythms with higher ingestion rates at night. The diel rhythms of C. typicus were maintained in the first laboratory-reared generation (F1), suggesting maternal effects, but disappeared in the following generations. The P. grani females of a long-term culture (> 10 years) also showed no differences in their day–night feeding activity. Ontogenetic variations were detected in the F1 generation of C. typicus: feeding rhythms were absent in naupliar stages, but adults fed more intensely at night. In the case of the cultured P. grani, in general none of the stages showed feeding rhythms. Laboratory-reared C. typicus (8–11 generations) did not recover back the natural cyclic feeding when exposed to jellyfish and fish exudates, indicating that either predation risk does not significantly affect their diel feeding activity or predator-induced responses in marine copepods might not involve chemical signalling. Our study confirms that feeding-related functional traits of marine copepods can experience quick multigenerational changes in the laboratory; consequently, cultured copepods might not be good models for studies involving their diel feeding behaviour.

Rapid shifts in the thermal sensitivity of growth but not development rate causes temperature–size response variability during ontogeny in arthropods

Size at maturity in ectotherms commonly declines with warming. This near‐universal phenomenon, formalised as the temperature–size rule, has been observed in over 80% of tested species, from bacteria to fish. The proximate cause has been attributed to the greater temperature dependence of development rate than growth rate, causing individuals to develop earlier but mature smaller in the warm. However, few studies have examined the ontogenetic progression of the temperature–size response at high resolution. Using marine planktonic copepods, we experimentally determined the progression of the temperature–size response over ontogeny. Temperature–size responses were not generated gradually from egg to adult, contrary to the predictions of a naïve model in which development rate was assumed to be more temperature‐dependent than growth rate, and the difference in the temperature dependence of these two rates remained constant over ontogeny. Instead, the ontogenetic progression of the temperature–size response in experimental animals was highly episodic, indicating rapid changes in the extent to which growth and development rates are thermally decoupled. The strongest temperature–size responses occurred temporally mid‐way through ontogeny, corresponding with the point at which individuals reached between ~5 and 25% of their adult mass. Using the copepod Oithona nana, we show that the temperature‐dependence of growth rate varied substantially throughout ontogeny, whereas the temperature dependence of development rate remained constant. The temperature‐dependence of growth rate even exceeded that of development rate in some life stages, leading to a weakening of the temperature–size response. Our analyses of arthropod temperature–size responses from the literature, including crustaceans and insects, support these conclusions more broadly. Overall, our findings provide a better understanding of how the temperature–size rule is produced over ontogeny. Whereas we find support for the generality of developmental rate isomorphy in arthropods (shared temperature dependence of development rate across life stages), this concept appears not to apply to growth rates.

Oxygen Consumption of Nauplii of Marine Planktonic Copepods

Oxygen Consumption of Nauplii of Marine Planktonic Copepods

Do functional groups of planktonic copepods differ in their ecological niches?

AbstractAimTo assess the degree of overlap between the environmental niches of marine planktonic copepods and test if the distribution of copepod functional groups differs across environmental gradients.LocationThe Mediterranean Sea.MethodsFunctional groups were defined based on clustering of functional traits in 106 marine copepod species using a multivariate ordination analysis. Functional traits included maximum body length, feeding mode, spawning strategy and trophic group. Simultaneously, the global distribution of the species was used to model their environmental niches with six environmental variables. For each of these predictors, four niche parameters were derived from the univariate response curve of each species to summarise their environmental preferences and ordinate the species in niche space through a PCA. Finally, the differences in the position in niche space of functional groups were tested with variance analysis.ResultsWe identified seven copepod functional groups with different distributions along the environmental gradients covered by our study. While carnivorous functional groups were affiliated with oligotrophic and tropical conditions, large and small current‐feeding herbivores are associated with colder, more seasonally varying and productive conditions. Small cruising detritivores and other small current‐feeding herbivores were not affiliated with specific conditions as their constituting species were scattered in niche space.Main conclusionsSince copepod functional groups occupy distinct ecological niches, ecosystem processes related to these groups are expected to vary across environmental gradients. Conditions favouring large current‐feeding herbivores should allow for enhanced fluxes of energy and nutrients through Mediterranean Sea ecosystems, while such fluxes should be weakened where large carnivores and small passive ambush‐feeding copepods dominate. Our study supports the development of trait‐based zooplankton functional groups in marine ecosystem models.

Effects of pH and temperature on egg hatching success of the marine planktonic copepod, Calanus finmarchicus

Calanus finmarchicus is a predominant planktonic copepod in the northern North Atlantic Ocean, where it is a fundamental link in the transfer of energy from phytoplankton to fish. Here, we investigate whether ocean acidification at present-day and future levels will cause a significant decrease in the egg hatching success (HS) of C. finmarchicus in the Gulf of Maine. Eggs spawned by female C. finmarchicus collected from the coastal Gulf of Maine were incubated in seawater acidified by addition of CO2 to selected pH levels at 3.5 °C (in a single experiment), 6 °C and 14–15 °C (in multiple experiments). HS was unaffected by pH between 6.58 and 8.0 at 3.5 and 6 °C, and between 7.1 and 8.0 when incubated at 15 °C. A significant interactive effect between temperature and pH on HS was found using a two-way ANOVA of the data from experiments at 6 °C and 14–15 °C, temperatures that are experienced in summer in the Gulf of Maine. HS of eggs spawned from C. finmarchicus females immediately after capture from a coastal station was significantly reduced at pH ≤ 7.0 when incubated at 14–15 °C, although HS of eggs collected from well-fed females in the laboratory in water from the Damariscotta Estuary was not significantly reduced at pH levels as low as 6.6 at 15 °C. This finding is consistent with the hypothesis that parental history and possibly maternal provisioning can influence capability of eggs to adjust to lower pH environments. While an interaction between pH and temperatures experienced by C. finmarchicus at the southern edge of its biogeographic range was observed, the pH at which this interaction occurred is substantially lower than pH levels predicted for the surface ocean over the surface ocean.

The complete mitochondrial genome of the copepod Calanus glacialis

Calanus glacialis, a marine planktonic copepod, is a keystone species in the Arctic Ocean. In this study, we shotgun sequenced the total DNA of one C. glacialis individual, using the NextSeq® Illumina platform, in order to determine its mitochondrial genome sequence. We successfully assembled and annotated this 20,674 bp long sequence, which included 13 protein-coding genes, 2 rRNA genes and 22 tRNA genes. Common gene-coding regions of 19 other species were used to reconstruct a phylogenetic tree, using mitogenomes of the phylogenetically closest copepods available. The new resource described here constitutes a tool of interest for better understanding the structure and dynamics of C. glacialis populations.

Magnetic light cloaking control in the marine planktonic copepod Sapphirina

We investigated the light cloaking behavior of the marine planktonic copepod Sapphirina under a magnetic field. Optical interferences in the multi-laminated guanine crystal layer beneath the dorsal body surface create a brilliant structural color, which can be almost entirely removed by changing the light reflection. In the investigation, we immersed segments of Sapphirina in seawater contained in an optical chamber. When the derived Sapphirina segments were attached to the container surface, they were inert to magnetic fields up to 300 mT. However, when the back plate segments were attached to the substrate at a point, with most of the plate floating in the seawater, the plate rotated oppositely to the applied magnetic field. In addition, the brilliant parts of the Sapphirina back plate rotated backward and forward by changing the magnetic field directions. Our experiment suggests a new model of an optical micro-electro-mechanical system that is controllable by magnetic fields.

Spliced leader-based analyses reveal the effects of polycyclic aromatic hydrocarbons on gene expression in the copepod Pseudodiaptomus poplesia

Polycyclic aromatic hydrocarbons (PAHs) are a group of toxic and carcinogenic pollutants that can adversely affect the development, growth and reproduction of marine organisms including copepods. However, knowledge on the molecular mechanisms regulating the response to PAH exposure in marine planktonic copepods is limited. In this study, we investigated the survival and gene expression of the calanoid copepod Pseudodiaptomus poplesia upon exposure to two PAHs, 1, 2-dimethylnaphthalene (1, 2-NAPH) and pyrene. Acute toxicity responses resulted in 96-h LC50 of 788.98μgL−1 and 54.68μgL−1 for 1, 2-NAPH and pyrene, respectively. Using the recently discovered copepod spliced leader as a primer, we constructed full-length cDNA libraries from copepods exposed to sublethal concentrations and revealed 289 unique genes of diverse functions, including stress response genes and novel genes previously undocumented for this species. Eighty-three gene families were specifically expressed in PAH exposure libraries. We further analyzed the expression of seven target genes by reverse transcription-quantitative PCR in a time-course test with three sublethal concentrations. These target genes have primary roles in detoxification, oxidative defense, and signal transduction, and include different forms of glutathione S-transferase (GST), glutathione peroxidases (GPX), peroxiredoxin (PRDX), methylmalonate-semialdehyde dehydrogenase (MSDH) and ras-related C3 botulinum toxin substrate (RAC1). Expression stability of seven candidate reference genes were evaluated and the two most stable ones (RPL15 and RPS20 for 1, 2-NAPH exposure, RPL15 and EF1D for pyrene exposure) were used to normalize the expression levels of the target genes. Significant upregulation was detected in GST-T, GST-DE, GPX4, PRDX6 and RAC1 upon 1, 2-NAPH exposure, and GST-DE and MSDH upon pyrene exposure. These results indicated that the oxidative stress was induced and that signal transduction might be affected by PAH exposure in P. poplesia. However, gene upregulation was followed by a reduction in expression level towards 96h, indicating a threshold value of exposure time that leads to depressed gene expression. Prolonged exposure may cause dysfunction of detoxification and antioxidant machinery in P. poplesia. The transcriptional responses of GST-T, GPX2 and GPX4 upon pyrene exposure were minimal. Our results reveal the different sensitivity of P. poplesia to two PAHs at both the individual and transcriptional levels. As the first attempt, this study proved that copepod spliced leader is useful for obtaining full-length cDNA in P. poplesia exposed to PAHs and provided a valuable gene resource for this non-model species. This approach can be applied to other calanoid copepods exposed to various stressors, particularly under field conditions.

Lethal and Sublethal Toxicity Comparison of BFRs to Three Marine Planktonic Copepods: Effects on Survival, Metabolism and Ingestion.

The estuarine planktonic copepods have a wide geographical distribution and commendable tolerance to various kinds of contaminants. The primary aim of the present study was to contrast the impacts of model POPs (TBBPA and HBCD) on three common estuarine planktonic copepods (Oithona similis, Acartia pacifica and Pseudodiaptomus inopinus) and establish a protocol for the assessment of acute toxicity of marine organic pollutants. We first quantified the 96h-LC50 (0.566, 0.04 and 0.257 mg/L of TBBPA to the three subjects above respectively and 0.314 mg/L of HBCD to P. inopinus; all reported concentrations are nominal values). In the sub-lethal toxicity tests, it was turned out that the effects of copepods exposed to TBBPA could product different influences on the energy ingestion and metabolism. Different type of pollutions, meanwhile, could also bring varying degree effect on the target copepods. In general, the indicators (the rate of oxygen consumption, ammonia excretion, food ingestion and filtration) in higher concentration groups showed marked significant difference compared with controls as well a dose-effect relationship. The study also extended the research on the joint toxicity of TBBPA and HBCD based on the survival rate of P.inopinus. Whether 1:1 concentration or 1:1 toxic level, the research showed synergy effect relative to single exposure conditions. The result indicated that current single ecological testing used for environmental protection activities may underestimate the risk for copepods. It was also demonstrated that short-term sub-lethal experiment could be a standard to evaluate the sensitivity of copepods to POPs.

Molecular Cloning of Secreted Luciferases from Marine Planktonic Copepods.

Secreted luciferases isolated from copepod crustaceans are frequently used for nondisruptive reporter-gene assays, such as the continuous, automated and/or high-throughput monitoring of gene expression in living cells. All known copepod luciferases share highly conserved amino acid residues in two similar, repeated domains in the sequence. The similarity in the domains are ideal nature for designing PCR primers to amplify cDNA fragments of unidentified copepod luciferases from bioluminescent copepod crustaceans. Here, we introduce how to establish a cDNA encoding novel copepod luciferases from a copepod specimen by PCR with degenerated primers.

How much crude oil can zooplankton ingest? Estimating the quantity of dispersed crude oil defecated by planktonic copepods

We investigated and quantified defecation rates of crude oil by 3 species of marine planktonic copepods (Temora turbinata, Acartia tonsa, and Parvocalanus crassirostris) and a natural copepod assemblage after exposure to mechanically or chemically dispersed crude oil. Between 88 and 100% of the analyzed fecal pellets from three species of copepods and a natural copepod assemblage exposed for 48 h to physically or chemically dispersed light crude oil contained crude oil droplets. Crude oil droplets inside fecal pellets were smaller (median diameter: 2.4–3.5 μm) than droplets in the physically and chemically dispersed oil emulsions (median diameter: 6.6 and 8.0 μm, respectively). This suggests that copepods can reject large crude oil droplets or that crude oil droplets are broken into smaller oil droplets before or during ingestion. Depending on the species and experimental treatments, crude oil defecation rates ranged from 5.3 to 245 ng-oil copepod−1 d−1, which represent a mean weight-specific defecation rate of 0.026 μg-oil μg-Ccopepod1 d−1. Considering a dispersed crude oil concentration commonly found in the water column after oil spills (1 μl L−1) and copepod abundances in high productive coastal areas, copepods may defecate ∼1.3–2.6 mg-oil m−3 d−1, which would represent ∼0.15%–0.30% of the total dispersed oil per day. Our results indicate that ingestion and subsequent defecation of crude oil by planktonic copepods has a small influence on the overall mass of oil spills in the short term, but may be quantitatively important in the flux of oil from surface water to sediments and in the transfer of low-solubility, toxic petroleum hydrocarbons into food webs after crude oil spills in the sea.

Ageing and Caloric Restriction in a Marine Planktonic Copepod.

Planktonic copepods are a key group in the marine pelagic ecosystem, linking primary production with upper trophic levels. Their abundance and population dynamics are constrained by the life history tradeoffs associated with resource availability, reproduction and predation pressure. The tradeoffs associated with the ageing process and its underlying biological mechanisms are, however, poorly known. Our study shows that ageing in copepods involves a deterioration of their vital rates and a rise in mortality associated with an increase in oxidative damage (lipid peroxidation); the activity of the cell-repair enzymatic machinery also increases with age. This increase in oxidative damage is associated with an increase in the relative content of the fatty acid 22:6(n-3), an essential component of cell membranes that increases their susceptibility to peroxidation. Moreover, we show that caloric (food) restriction in marine copepods reduces their age-specific mortality rates, and extends the lifespan of females and their reproductive period. Given the overall low production of the oceans, this can be a strategy, at least in certain copepod species, to enhance their chances to reproduce in a nutritionally dilute, temporally and spatially patchy environment.

Mandibular gnathobases of marine planktonic copepods - feeding tools with complex micro- and nanoscale composite architectures.

Copepods are dominant members of the marine zooplankton. Their diets often comprise large proportions of diatom taxa whose silicified frustules are mechanically stable and offer protection against grazers. Despite of this protection, many copepod species are able to efficiently break even the most stable frustule types. This ability requires specific feeding tools with mechanically adapted architectures, compositions and properties. When ingesting food, the copepods use the gnathobases of their mandibles to grab and, if necessary, crush and mince the food items. The morphology of these gnathobases is related to the diets of the copepods. Gnathobases of copepod species that mainly feed on phytoplankton feature compact and stable tooth-like structures, so-called teeth. In several copepod species these gnathobase teeth have been found to contain silica. Recent studies revealed that the siliceous teeth are complex microscale composites with silica-containing cap-like structures located on chitinous exoskeleton sockets that are connected with rubber-like bearings formed by structures with high proportions of the soft and elastic protein resilin. In addition, the silica-containing cap-like structures exhibit a nanoscale composite architecture. They contain some amorphous silica and large proportions of the crystalline silica type α-cristobalite and are pervaded by a fine chitinous fibre network that very likely serves as a scaffold during the silicification process. All these intricate composite structures are assumed to be the result of a coevolution between the copepod gnathobases and diatom frustules in an evolutionary arms race. The composites very likely increase both the performance of the siliceous teeth and their resistance to mechanical damage, and it is conceivable that their development has favoured the copepods’ dominance of the marine zooplankton observed today.

Hydrostatic pressure and temperature effects on the membranes of a seasonally migrating marine copepod.

Marine planktonic copepods of the order Calanoida are central to the ecology and productivity of high latitude ecosystems, representing the interface between primary producers and fish. These animals typically undertake a seasonal vertical migration into the deep sea, where they remain dormant for periods of between three and nine months. Descending copepods are subject to low temperatures and increased hydrostatic pressures. Nothing is known about how these organisms adapt their membranes to these environmental stressors. We collected copepods (Calanoides acutus) from the Southern Ocean at depth horizons ranging from surface waters down to 1000 m. Temperature and/or pressure both had significant, additive effects on the overall composition of the membrane phospholipid fatty acids (PLFAs) in C. acutus. The most prominent constituent of the PLFAs, the polyunsaturated fatty acid docosahexanoic acid [DHA – 22:6(n-3)], was affected by a significant interaction between temperature and pressure. This moiety increased with pressure, with the rate of increase being greater at colder temperatures. We suggest that DHA is key to the physiological adaptations of vertically migrating zooplankton, most likely because the biophysical properties of this compound are suited to maintaining membrane order in the cold, high pressure conditions that persist in the deep sea. As copepods cannot synthesise DHA and do not feed during dormancy, sufficient DHA must be accumulated through ingestion before migration is initiated. Climate-driven changes in the timing and abundance of the flagellated microplankton that supply DHA to copepods have major implications for the capacity of these animals to undertake their seasonal life cycle successfully.

Infection of the planktonic copepod Calanus finmarchicus by the parasitic dinoflagellate, Blastodinium spp: effects on grazing, respiration, fecundity and fecal pellet production

Several dinoflagellate species in the genus Blastodinium are gut parasites of marine planktonic copepods. However,there is only limited information on the occurrence and infection frequencies of Blastodinium spp. in the field andalmost no information on the functional impact on their hosts. We report upon the effects of Blastodinium sp. infectionon Calanus finmarchicus from the northeastern Atlantic coast off southern Norway during April 2013 and 2014. Up to58% of C. finmarchicus were infected near the coast, while ,5% were infected several kilometers offshore. Ingestionrates of infected females were below detection limits and significantly lower than uninfected females. Blastodinium sp.-infected females showed characteristic symptoms of starvation, including lower respiration rates (implying a lowermetabolic rate), production of smaller and fewer fecal pellets and significantly fewer eggs than uninfected females. Afew females in this study were able to void the infection, however the extended period of starvation is likely to havelonger-term repercussions on egg production rates well after the copepod clears the infection. The degree to whichthe infection affects C. finmarchicus recruitment depends on the extent of the spatial distribution of the infection.Monitoring of parasitic infection during routine field surveys will be required in order to clarify this.KEYWORDS: zooplankton; secondary production; gut parasite; starvation; egg production; sterility; metabolism;ingestion; population dynamics; biogeochemical cycles

A metagenetic approach for revealing community structure of marine planktonic copepods.

Marine planktonic copepods are an ecologically important group with high species richness and abundance. Here, we propose a new metagenetic approach for revealing the community structure of marine planktonic copepods using 454 pyrosequencing of nuclear large subunit ribosomal DNA. We determined an appropriate similarity threshold for clustering pyrosequencing data into molecular operational taxonomic units (MOTUs) using an artificial community containing 33 morphologically identified species. The 99% similarity threshold had high species-level resolution for MOTU clustering but overestimated species richness. The artificial community was appropriately clustered into MOTUs at 97% similarity, with little inflation in MOTU numbers and with relatively high species-level resolution. The number of sequence reads of each MOTU was correlated with dry weight of that taxon, suggesting that sequence reads could be used as a proxy for biomass. Next, we applied the method to field-collected samples, and the results corresponded reasonably well with morphological analysis of these communities. Numbers of MOTUs were well correlated with species richness at 97% similarity, and large numbers of sequence reads were generally observed in MOTUs derived from species with large biomass. Further, MOTUs were successfully classified into taxonomic groups at the family level at 97% similarity; similar patterns of species richness and biomass were revealed within families with metagenetic and morphological analyses. At the 99% similarity threshold, MOTUs with high proportions of sequence reads were identified as biomass-dominant species in each field-collected sample. The metagenetic approach reported here can be an effective tool for rapid and comprehensive assessment of copepod community structure.