Oxidation kinetics and inverse isotope effect of marine nitrite-oxidizing isolates

Nitrification, the step-wise oxidation of ammonium to nitrite and nitrate, is important in the marine environment because it produces nitrate, the most abundant marine dissolved inorganic nitrogen (DIN) component and N-source for phytoplankton and microbes. This study focused on the second step of nitrification, which is carried out by a distinct group of organisms, nitrite-oxidizing bacteria (NOB). The growth of NOB is characterized by nitrite oxidation kinetics, which we investigated for 4 pure cultures of marine NOB (Nitrospina watsonii 347, Nitrospira sp. Ecomares 2.1, Nitrococcus mobilis 231, and Nitrobacter sp. 311). We further compared the kinetics to those of non-marine species because substrate concentrations in marine environments are comparatively low, which likely influences kinetics and highlights the importance of this study. We also determined the isotope effect during nitrite oxidation of a pure culture of Nitrospina (Nitrospina watsonii 347) belonging to one of the most abundant marine NOB genera, and for a Nitrospira strain (Nitrospira sp. Ecomares 2.1). The enzyme kinetics of nitrite oxidation, described by Michaelis-Menten kinetics, of 4 marine genera are rather narrow and fall in the low end of half-saturation constant (Km) values reported so far, which span over 3 orders of magnitude between 9 and >1000 µM NO2-. Nitrospina has the lowest Km (19 µM NO2-), followed by Nitrobacter (28 µM NO2-), Nitrospira (54 µM NO2-), and Nitrococcus (120 µM NO2-). The isotope effects during nitrite oxidation by Nitrospina watsonii 347 and Nitrospira sp. Ecomares 2.1 were 9.7 ± 0.8 and 10.2 ± 0.9‰, respectively. This confirms the inverse isotope effect of NOB described in other studies; however, it is at the lower end of reported isotope effects. We speculate that differences in isotope effects reflect distinct nitrite oxidoreductase (NXR) enzyme orientations

Diet and stable isotope analyses reveal the feeding ecology of the orangeback squid Sthenoteuthis pteropus (Steenstrup 1855) (Mollusca, Ommastrephidae) in the eastern tropical Atlantic

In the eastern tropical Atlantic, the orangeback flying squid Sthenoteuthis pteropus (Steenstrup 1855) (Cephalopoda, Ommastrephidae) is a dominant species of the epipelagic nekton community. This carnivore squid has a short lifespan and is one of the fastest-growing squids. In this study, we characterise the role of S. pteropus in the pelagic food web of the eastern tropical Atlantic by investigating its diet and the dynamics of its feeding habits throughout its ontogeny and migration. During three expeditions in the eastern tropical Atlantic in 2015, 129 specimens were caught by hand jigging. Stomach content analyses (via visual identification and DNA barcoding) were combined with stable isotope data (∂15N and ∂13C) of muscle tissue to describe diet, feeding habits and trophic ecology of S. pteropus. Additionally, stable isotope analyses of incremental samples along the squid’s gladius—the chitinous spiniform structure supporting the muscles and organs—were carried out to explore possible diet shifts through ontogeny and migration. Our results show that S. pteropus preys mainly on myctophid fishes (e.g. Myctophum asperum, Myctophum nitidulum, Vinciguerria spp.), but also on other teleost species, cephalopods (e.g. Enoploteuthidae, Bolitinidae, Ommastrephidae), crustaceans and possibly on gelatinous zooplankton as well. The squid shows a highly opportunistic feeding behaviour that includes cannibalism. Our study indicates that the trophic position of S. pteropus may increase by approximately one trophic level from a mantle length of 15 cm to 47 cm. The reconstructed isotope-based feeding chronologies of the gladii revealed high intra- and inter-individual variability in the squid’s trophic position and foraging area. These findings are not revealed by diet or muscle tissue stable isotope analysis. This suggests a variable and complex life history involving individual variation and migration. The role of S. pteropus in transferring energy and nutrients from lower to higher trophic levels may be underestimated and important for understanding how a changing ocean impacts food webs in the eastern Atlantic

A novel metabarcoding primer pair for environmental DNA analysis of Cephalopoda (Mollusca) targeting the nuclear 18S rRNA region

Cephalopods are pivotal components of marine food webs, but biodiversity studies are hampered by challenges to sample these agile marine molluscs. Metabarcoding of environmental DNA (eDNA) is a potentially powerful technique to study oceanic cephalopod biodiversity and distribution but has not been applied thus far. We present a novel universal primer pair for metabarcoding cephalopods from eDNA, Ceph18S (Forward: 5′-CGC GGC GCT ACA TAT TAG AC-3′, Reverse: 5′-GCA CTT AAC CGA CCG TCG AC-3′). The primer pair targets the hypervariable region V2 of the nuclear 18S rRNA gene and amplifies a relatively short target sequence of approximately 200 bp in order to allow the amplification of degraded DNA. In silico tests on a reference database and empirical tests on DNA extracts from cephalopod tissue estimate that 44-66% of cephalopod species, corresponding to about 310-460 species, can be amplified and identified with this primer pair. A multi-marker approach with the novel Ceph18S and two previously published cephalopod mitochondrial 16S rRNA primer sets targeting the same region (Jarman et al. 2006 Mol. Ecol. Notes. 6, 268-271; Peters et al. 2015 Mar. Ecol. 36, 1428-1439) is estimated to amplify and identify 89% of all cephalopod species, of which an estimated 19% can only be identified by Ceph18S. All sequences obtained with Ceph18S were submitted to GenBank, resulting in new 18S rRNA sequences for 13 cephalopod tax

The role of gelatinous macrozooplankton in deep-sea carbon transport in Cape Verde Cruise No. POS532

4/2/2019 – 24/2/2019, Mindelo (Republic of Cape Verde) – Mindelo (Republic of Cape Verde) DeepC-Jelly We proposed to test the hypothesis that large gelatinous macrozooplankton (e.g. tunicates, hydrozoans) are a significant carbon storage in midwater, and a vector for carbon from midwater to the ocean floor in Cape Verde. To test this hypothesis, we studied 1) the distribution, diversity and abundance of gelatinous organisms in the epi-, meso-, and bathypelagic zone, 2) their role in transporting carbon through the pelagic foodweb to the seafloor and 3) their behavior and associations. We worked in the coastal deep sea off Santo Antão and Fogo as well as in the open ocean at the time series station CVOO and an eddy. A manned submersible was used for mesopelagic surveys, to document the behaviour and associations of deep-sea organisms and to collect living specimens. We performed pelagic video transects, discrete net sampling, and eDNA sampling down to 3000 m. ADCP and CTD transects allowed a detailed reconstruction of the effect of the islands on currents and productivity. To quantify the carbon flux of pelagic foodfalls, we also surveyed the seafloor. Sample and video analysis is still in progress, but first results indicate the impact of the pelagic tunicate Pyrosoma atlanticum, which is an upwelling-favored species largely absent from the oligotrophic open ocean, in the nearshore regions of Cape Verde as well as in the cyclonic eddy sampled. It was also observed on the seafloor and resembles a food source and a habitat in the water column and in the benthos. Specimens of pelagic fauna were collected that allow new species descriptions. New records and a new species for the region were also observed during benthic surveys. The cruise was documented in various outreach activities including national television in Cabo Verde

Arctic Seafloor Integrity Cruise No. MSM95 – (GPF 19-2_05)

The main aim of the MSM95 research expedition was to investigate and map physical impacts on the arctic seafloor in two distinct and contrasting Arctic areas (The Svalbard shelf edge and the HAUSGARTEN time series stations in the FRAM strait) with a range of research equipment. A ‘nested’ data approach was conducted in each research area, with broad seafloor mapping conducted initially with the R/V MARIA S. MERIAN onboard acoustic systems (The EM122 and EM712 bathymetric systems), followed by focused subsequent mapping conducted by PAUL 3000 automated underwater vehicle (AUV) sidescan and camera deployments, Ocean Floor Observation and Bathymetry System (OFOBS) towed sidescan and camera trawls and finally with very high resolution investigations conducted with a new mini-ROV launched directly from the OFOBS for close seafloor visual analysis. These data will be used to produce spatial distribution maps of iceberg and fishery impacts on the seafloor at three locations to the north, south and west of the Svalbard Archipelago, as well as maps of drop stone and topography variations across several of the HAUSGARTEN stations

Deep-sea predator niche segregation revealed by combined cetacean biologging and eDNA analysis of cephalopod prey

Fundamental insight on predator-prey dynamics in the deep sea is hampered by a lack of combined data on hunting behavior and prey spectra. Deep-sea niche segregation may evolve when predators target specific prey communities, but this hypothesis remains untested. We combined environmental DNA (eDNA) metabarcoding with biologging to assess cephalopod community composition in the deep-sea foraging habitat of two top predator cetaceans. Risso’s dolphin and Cuvier’s beaked whale selectively targeted distinct epi/meso- and bathypelagic foraging zones, holding eDNA of 39 cephalopod taxa, including 22 known prey. Contrary to expectation, extensive taxonomic overlap in prey spectra between foraging zones indicated that predator niche segregation was not driven by prey community composition alone. Instead, intraspecific prey spectrum differences may drive differentiation for hunting fewer, more calorific, mature cephalopods in deeper waters. The novel combination of methods presented here holds great promise to disclose elusive deep-sea predator-prey systems, aiding in their protection