Description: Simultaneous analysis of carbon, nitrogen and sulphur stable isotope ratios was applied in this pilot study to examine the food web of a Zostera marina L. system in the western Baltic Sea. Samples of three potential food sources: eelgrass, epiphytic algae and seston, as well as 69 consumer species were collected during the growing season of Z. marina from March to September 2011. The measured δ13C values of epiphytes (-14.1‰ ± 1.8 SD) were close to δ13C values of eelgrass (-11.6‰ ± 1.8 SD), impeding a clear distinction of those two carbon sources, whereas seston δ13C values (-20.9‰ ± 3.5 SD) were clearly different. This frequently encountered problem was solved by the additional use of δ34S, which resulted in easily distinguishable values for sediment and seawater derived sulphur. Values of primary producer δ34S ranged from 5.6‰ (± 2.3 SD) for Z. marina leaves to 14.2‰ (± 1.6 SD) for epiphytes and 11.9‰ (± 3.3 SD) for seston. The combination of δ34S and δ13C values made a separation of carbon sources possible and enabled the allocation of potential food sources to consumers and a description of their trophic relationships. The data of stable isotope ratio analysis of this eelgrass community strongly indicate a food web based on epiphyte and seston production. δ15N values show a food web consisting of large numbers of generalists and a high degree of omnivory amongst the consumer species analysed. This implies an occupation of every trophic position possible, which is supported by a continuous distribution of δ15N values. Previously described eelgrass food webs may have to be re-evaluated to include sulfur in order to provide a clear picture on primary carbon sources.

Description: The differences in the impact of two major groups of herbivorous zooplankton (Cladocera and Copepoda) on summer phytoplankton in a mesotrophic lake were studied. Field experiments were performed in which phytoplankton were exposed to different densities of two major types of herbivorous zooplankton, cladocerans and copepods. Contrary to expectation, neither of the two zooplankton groups significantly reduced phytoplankton biomass. However, there were strong and contrasting impacts on phytoplankton size structure and on individual taxa. Cladocerans suppressed small phytoplankton, while copepods suppressed large phytoplankton. The unaffected size classes compensated for the loss of those affected by enhanced growth. After contamination of the copepod mesocosms with the cladoceran Daphnia, the combined impact of both zooplankton groups caused a decline in total phytoplankton biomass.

Description: The foraging modes of calanoid copepods differ in that stationary suspension-feeding is more easily detected by prey with strong escape responses (ciliates) than is ‘cruising’ or ‘ambushing’ feeding. Thus, the ability of a copepod to include heterotrophic prey in its diet may be associated with its foraging mode and, further, with its nitrogen stable isotope signature (δ15N). This is because a more carnivorous diet may be expected to result in a higher δ15N. We tested this hypothesis in a mesocosm study using a density gradient (0 to 80 ind. l-1) of calanoid copepods. We expected copepod δ15N to generally increase with decreasing copepod density because of increased food availability, and predicted stronger increases for cruising than for stationary suspension-feeding species. As an assemblage, copepods had a pronounced impact on the food web: diatoms and ciliates decreased, whereas nanoflagellates increased with increasing copepod density. As expected, Centropages hamatus, a cruising species, showed the strongest isotopic increase and also highest population growth at low copepod density, suggesting that it was the most efficient species in capturing ciliates. Temora longicornis, a stationary suspension-feeder, showed a uniform isotopic increase in all mesocosms, which we believe resulted from nutritional stress arising from poor feeding on both ciliates (too fast for ingestion by T. longicornis) and nanoflagellates (too small). However, Pseudocalanus elongatus, a species equally categorised as a stationary suspension-feeder, showed increases in its δ15N similar to those for C. hamatus. While this may indicate potential switching in its foraging mode, alternative explanations cannot be ruled out, partly because qualitative and quantitative aspects of trophic enrichment in our experiment could not be clearly separated. This study shows that consumer δ15N are difficult to interpret, even if potential food sources and aspects of the species’ biology are known, and thus emphasises the necessity for further laboratory studies to help better interpret zooplankton δ15N in the field.

Description: Zooplankton grazing on bacterio- and phytoplankton was studied in the Gulf of Aqaba and the Northern Red Sea during Meteor Cruise Me 44-2 in February-March 1999. Protozoan grazing on bacterioplankton and autotrophic ultraplankton was studied by the Landry dilution method. Microzooplankton grazing on phytoplankton 〉6 µm was studied by incubation experiments in the presence and absence of microzooplankton. Mesozooplankton grazing was studied by measuring per capita clearance rates of individual zooplankton with radioactively labelled food organisms and estimating in situ rates from abundance values. Protozoan grazing rates on heterotrophic bacteria and on algae 〈6 µm were high (bacteria: 0.7 to 1.1 d-1, ultraphytoplankton: 0.7 to 1.3 d-1), while grazing rates on Synechococcus spp. were surprisingly low and undetectable in some experiments. Mesozooplankton grazing was weak, cumulative grazing rates being ca. 2 orders of magnitude smaller than the grazing rates by protozoans. Among mesozooplankton, appendicularians specialised on smaller food items and calanoid copepods on larger ones.

Description: A common elemental analyzer system connected to a temperature-controlled gas chromatography (GC) column and coupled to an isotope ratio mass spectrometer was improved to decrease the determination limit for a simultaneous stable isotope ratio measurement of nitrogen and carbon dioxide. The additional use of a special ashtray system to collect the combustion residuals permitted more time-efficient work. These modifications to the elemental analyzer allowed precise measurements to be made down to 1.5 µg nitrogen and 10 µg carbon for stable isotope analysis. Low system background values and an acceptable signal-to-noise ratio have made an additional blank correction for these low sample measurements unnecessary. We provide a precision of this stable isotope analysis for lowest amounts of 1.2–2 µg nitrogen with a standard deviation of ±0.496‰ (n = 27) and for 8.2–15 µg carbon with a standard deviation of ±0.257‰ (n = 31) across different sample runs under stipulated conditions. This application can be established in an automatic mode without cryofocusing procedures.

Description: Marine food webs are highly compartmentalized, and characterizing the trophic niches among consumers is important for predicting how impact from human activities affects the structuring and functioning of marine food webs. Biomarkers such as bulk stable isotopes have proven to be powerful tools to elucidate trophic niches, but they may lack in resolution, particularly when spatiotemporal variability in a system is high. To close this gap, we investigated whether carbon isotope (delta C-13) patterns of essential amino acids (EAAs), also termed delta(13)C(AA)fingerprints, can characterize niche differentiation in a highly dynamic marine system. Specifically, we tested the ability of delta(13)C(AA)fingerprints to differentiate trophic niches among six functional groups and ten individual species in the Baltic Sea. We also tested whether fingerprints of the common zooplanktivorous fishes, herring and sprat, differ among four Baltic Sea regions with different biochemical conditions and phytoplankton assemblages. Additionally, we investigated how these results compared to bulk C and N isotope data for the same sample set. We found significantly different delta(13)C(AA)fingerprints among all six functional groups. Species differentiation was in comparison less distinct, due to partial convergence of the species' fingerprints within functional groups. Herring and sprat displayed region-specific delta(13)C(AA)fingerprints indicating that this approach could be used as a migratory marker. Niche metrics analyses showed that bulk isotope data had a lower power to differentiate between trophic niches than delta(13)C(AA)fingerprinting. We conclude that delta(13)C(AA)fingerprinting has a strong potential to advance our understanding of ecological niches, and trophic linkages from producers to higher trophic levels in dynamic marine systems. Given how management practices of marine resources and habitats are reshaping the structure and function of marine food webs, implementing new and powerful tracer methods are urgently needed to improve the knowledge base for policy makers.

Description: Mixotrophic organisms are increasingly recognized as important components of ecosystems, but the factors controlling their nutrition pathways (in particular their autotrophy-heterotrophy balance) are little known. Both autotrophy and heterotrophy are expected to respond to density-dependent mechanisms but not necessarily in the same direction and/or strength. We hypothesize that the autotrophy-heterotrophy balance of mixotrophic organisms might therefore be a function of population densities. To investigate this relationship, we sampled mixotrophic jellyfish holobionts (host, Mastigias papua etpisoni; symbiont, Cladocopium sp.) in a marine lake (Palau, Micronesia) on six occasions (from 2010 to 2018). Over this period, population densities varied similar to 100 fold. We characterized the nutrition of the holobionts using the delta C-13 and delta N-15 signatures as well as C:N ratios. delta C-13 values increased and delta N-15 values decreased with increasing population densities (respectively, R-2 = 0.86 and 0.70, P 〈 0.05). Although less distinct, C:N ratios increased with increasing population densities (R-2 = 0.59, 0.1 〉 P 〉 0.05). This indicates that the autotrophy-heterotrophy balance tends toward autotrophy when population densities increase. We propose that the availability of zooplanktonic prey is the main driver of this pattern. These results demonstrate that the autotrophy-heterotrophy balance of mixotrophic jellyfishes can be tightly regulated by density-dependent mechanisms.

Description: We fertilised 12 mesocosms with NE Atlantic phytoplankton with different Si:N ratios (0:1 to 1:1). After 1 wk, we added mesozooplankton, mainly calanoid copepods at natural densities to 10 of the mesocosms; the remaining 2 mesocosms served as controls. A trend of increasing diatom dominance with increasing Si:N ratios and species-specific correlations of diatoms to Si:N ratios were not changed by the addition of mesozooplankton. Large unicellular and chain-forming diatoms, thin-walled dinoflagellates (Gymnodiniales) and ciliates were reduced by copepod grazing while armoured dinoflagellates remained unaffected. Nanoplanktonic flagellates and diatoms profited from the addition of copepods, probably through release from ciliate grazing.

Description: In a mesocosm experiment in Kiel Fjord, the plankton community 〈250 µm was exposed to a mesozooplankton density gradient (5 to 80 individuals l–1) dominated by the calanoid copepod Acartia clausi. Over the experimental period (9 d), the diazotrophic cyanobacterium Nodularia spumigena increased exponentially, irrespective of mesozooplankton densities, attaining maximum concentrations of 1200 cells ml–1 (3700 filaments l–1). At the end of the experiment, the δ15N of particulate organic matter was negatively correlated with N. spumigena concentrations, indicating the fixation of isotopically ‘light’, diazotrophic nitrogen. In all treatments, final copepod δ15N were lower (–0.7 to –2.7‰) than initial copepod δ15N, indicating the transfer of diazotrophic nitrogen to mesozooplankton. Based on a simple isotopic mixing model, diazotrophic nitrogen was calculated to contribute 45 to 6% to final copepod δ15N along the mesozooplankton gradient. This translates to a transfer of 2 to 24% of net nitrogen fixation to the mesozooplankton standing stock. The absence of any mesozooplankton impact on N. spumigena, and the negative impact found for other microplankton, including diatoms and ciliates, suggest that diazotrophically fixed nitrogen reached mesozooplankton indirectly through trophic vectors. This is consistent with the fact that copepod δ15N decreased with decreasing mesozooplankton densities, since only a quantitatively limited dietary source may be expected to result in density-dependent changes in copepod δ15N. Considering that natural mesozooplankton densities in the Baltic Sea rarely exceed 10 ind. l–1, the contribution of diazotrophically fixed nitrogen to mesozooplankton may be substantial (23 to 45%) during summer blooms of diazotrophic cyanobacteria.

Description: A recent meta-analysis indicates that trophic cascades (indirect effects of predators on plants via herbivores) are weak in marine plankton in striking contrast to freshwater plankton (Shurin et al. 2002, Ecol. Lett., 5, 785–791). Here we show that in a marine plankton community consisting of jellyfish, calanoid copepods and algae, jellyfish predation consistently reduced copepods but produced two distinct, opposite responses of algal biomass. Calanoid copepods act as a switch between alternative trophic cascades along food chains of different length and with counteracting effects on algal biomass. Copepods reduced large algae but simultaneously promoted small algae by feeding on ciliates. The net effect of jellyfish on total algal biomass was positive when large algae were initially abundant in the phytoplankton, negative when small algae were dominant, but zero when experiments were analysed in combination. In contrast to marine systems, major pathways of energy flow in Daphnia-dominated freshwater systems are of similar chain length. Thus, differences in the length of alternative, parallel food chains may explain the apparent discrepancy in trophic cascade strength between freshwater and marine planktonic systems.