Benthic foraminiferal responses to anthropogenic manipulation on a wild oyster reef

Benthic foraminifera are known to respond quickly to environmental changes. Biological indices are used in marine subtidal areas (Barras et al., 2014) to monitor impacted marine environments (Mojtahid et al., 2006, 2008). Such indices could be useful in transitional marine environments such as estuaries and mudflats where no accurate indices are available. To develop such to ols and to improve our knowledge on b enthic foraminifera in intertidal environments, we followed the spatial and temporal distribution of b enthic foraminifera in Bourgneuf Bay, a coastal bay with a large mudflat situated south of the Loire estuary on French west coast. At this location, wild oyster reefs are present. Partial release of organic and inorganic nutrients through oyster excretion enriches the nutrients flow leading to microphytobenthos bloom around the reef. To show this relation and to understand the relation between the different biological compartments,we analysed the foraminiferal composition of the sediment around the reef before and after ananthrop ogenic manipulation. In July 2014, the oysters of the reef were removed in order to assess their impact on microphytob enthos and meiofauna. In this study, we showed a strong impact of this anthrop ogenic manipulation on benthic foraminiferal densities and species composition. These results confirmed the strong and quick responses of benthic foraminifera from intertidal areas to environmental changes and showed that intertidal benthic foraminifera can be used as an index to monitor transitional marine environments

Locomotion speed of the benthic foraminifer Ammonia tepida exposed to different nitrogen and carbon sources

Ammonia tepida is a dominant benthic foraminifer colonizing intertidal mudflat sediments. Horizontal locomotion speeds were monitored using time-lapse image analysis over 6 and 24 h. Experimental conditions were based on foraminifera exposed to dry sediment re-suspended in artificial sea water (ASW) without any nutrient addition (condition DS), to combusted sediment re-suspended in in ASW also without any nutrient addition (condition CS), or to combusted sediment re-suspended in ASW enriched with either: nitrate, urea, glucose, soil extract (SE), extracellular polymeric substances (EPS), benthic diatoms (Entomoneis paludosa) or natural microphytobenthic assemblages (MPB). Significant differences were already measured after 6 h between A. tepida mean locomotion speeds at the different experimental conditions. However, differences were clearer after 24 h where the slowest A. tepida mean locomotion speed was measured in specimens placed in CS (1.00 ± 0.30 mm h− 1) and the highest mean locomotion speed in DS (2.99 ± 0.22 mm h− 1). Three different groups were defined according to their locomotion speed, (1) foraminifera exposed to DS had a locomotion speed significantly higher than all other conditions, (2) foraminifera placed in conditions enriched in SE, Glucose, Urea and EPS had intermediary locomotion speeds (1.8–2.5 mm h− 1), and (3) conditions with foraminifera showing the lowest locomotion speeds (1–1.6 mm h− 1) were CS, nitrate, MPB and E. paludosa. Thus, foraminifera exposed to organic matter (DS, SE, Glucose and Urea) showed faster locomotion speeds than foraminifera exposed to inorganic matter (CS, nitrate) or live preys (E. paludosa, MPB). Dissolved organic matter enrichment enhanced foraminifera locomotion speed, which might be a behavioural response to satisfy their carbon and/or nitrogen requirements, and the lowest locomotion speed observed when feeding on live preys might be a consequence of longer time required for live prey phagocytosis

What happened to my plastids? - Persistence of functionality in diatom plastids stolen by grazer intertidal benthic foraminifera

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Inorganic carbon and nitrogen assimilation in cellular compartments of a benthic kleptoplastic foraminifer

© The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Scientific Reports 8 (2018): 10140, doi:10.1038/s41598-018-28455-1.Haynesina germanica, an ubiquitous benthic foraminifer in intertidal mudflats, has the remarkable ability to isolate, sequester, and use chloroplasts from microalgae. The photosynthetic functionality of these kleptoplasts has been demonstrated by measuring photosystem II quantum efficiency and O2 production rates, but the precise role of the kleptoplasts in foraminiferal metabolism is poorly understood. Thus, the mechanism and dynamics of C and N assimilation and translocation from the kleptoplasts to the foraminiferal host requires study. The objective of this study was to investigate, using correlated TEM and NanoSIMS imaging, the assimilation of inorganic C and N (here ammonium, NH4+) in individuals of a kleptoplastic benthic foraminiferal species. H. germanica specimens were incubated for 20 h in artificial seawater enriched with H13CO3− and 15NH4+ during a light/dark cycle. All specimens (n = 12) incorporated 13C into their endoplasm stored primarily in the form of lipid droplets. A control incubation in darkness resulted in no 13C-uptake, strongly suggesting that photosynthesis is the process dominating inorganic C assimilation. Ammonium assimilation was observed both with and without light, with diffuse 15N-enrichment throughout the cytoplasm and distinct 15N-hotspots in fibrillar vesicles, electron-opaque bodies, tubulin paracrystals, bacterial associates, and, rarely and at moderate levels, in kleptoplasts. The latter observation might indicate that the kleptoplasts are involved in N assimilation. However, the higher N assimilation observed in the foraminiferal endoplasm incubated without light suggests that another cytoplasmic pathway is dominant, at least in darkness. This study clearly shows the advantage provided by the kleptoplasts as an additional source of carbon and provides observations of ammonium uptake by the foraminiferal cell.This work was supported by the Swiss National Science Foundation (grant no. 200021_149333) and was part of the CNRS EC2CO-Lefe project ForChlo. It was also supported by the Region Pays de la Loire (Post-doc position of TJ, on FRESCO project) as well as the WHOI Robert W. Morse Chair for Excellence in Oceanography and The Investment in Science Fund at WHOI

Ultrastructure and distribution of kleptoplasts in benthic foraminifera from shallow-water (photic) habitats

© The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Marine Micropaleontology 138 (2018): 46-62, doi:10.1016/j.marmicro.2017.10.003.Assimilation, sequestration and maintenance of foreign chloroplasts inside an organism is termed “chloroplast sequestration” or “kleptoplasty”. This phenomenon is known in certain benthic foraminifera, in which such kleptoplasts can be found both intact and functional, but with different retention times depending on foraminiferal species. In the present study, seven species of benthic foraminifera (Haynesina germanica, Elphidium williamsoni, E. selseyense, E. oceanense, E. aff. E. crispum, Planoglabratella opercularis and Ammonia sp.) were collected from shallow-water benthic habitats and examined with transmission electron microscope (TEM) for cellular ultrastructure to ascertain attributes of kleptoplasts. Results indicate that all these foraminiferal taxa actively obtain kleptoplasts but organized them differently within their endoplasm. In some species, the kleptoplasts were evenly distributed throughout the endoplasm (e.g., H. germanica, E. oceanense, Ammonia sp.), whereas other species consistently had plastids distributed close to the external cell membrane (e.g., Elphidium williamsoni, E. selseyense, P. opercularis). Chloroplast degradation also seemed to differ between species, as many degraded plastids were found in Ammonia sp. and E. oceanense compared to other investigated species. Digestion ability, along with different feeding and sequestration strategies may explain the differences in retention time between taxa. Additionally, the organization of the sequestered plastids within the endoplasm may also suggest behavioral strategies to expose and/or protect the sequestered plastids to/from light and/or to favor gas and/or nutrient exchange with their surrounding habitats.TJ was funded by the “FRESCO” project, a project supported by the Region Pays de Loire and the University of Angers. This work was also supported by a grant no. 200021_149333 from the Swiss National Science Foundation and the French national program EC2CO-LEFE (project ForChlo).JMB acknowledges the Robert W. Morse Chair for Excellence in Oceanography and the Investment in Science Fund at WHOI. Also, KK acknowledges the Academy of Finland (Project numbers: 278827, 283453)

Effect of bacteria on growth and biochemical composition of two benthic diatoms Halamphora coffeaeformis and Entomoneis paludosa

Benthic diatoms are the dominant microalgae in intertidal mudflats and are in constant interaction with their surrounding bacteria. This study was designed to investigate the effect of bacteria on growth, biomass, elemental (C & N) and biochemical composition, and extracellular polymeric substances (EPS) excretion by two marine benthic diatoms, Halamphora coffeaeformis and Entomoneis paludosa. The experiments were conducted on diatom cultures previously exposed or not to antibiotics. The treatment with antibiotics caused a decrease of bacterial abundance from 24 to fewer than 1 bacteria per algal cell. In non-treated cultures of E. paludosa and H. coffeaeformis, the bacteria phylogenetic affiliation was equally distributed between Bacteroidetes (Flavobacteriia) and Proteobacteria (alpha- and gammaproteobacteria). After treatment with antibiotics, the residual bacterial community was ~ 37% Flavobacteriia (Winogragskyella genus), 34% for the alphaproteobacteria (mainly Roseibacterium sp and Antarctobacter sp.) and 29% for the gammaproteobacteria (mainly Methylophaga sp. and Stenotrophomonas sp.). Growth of H. coffeaeformis and E. paludosa in non-treated cultures was enhanced by the abundance of the associated bacteria, with mean growth rate of 1 day− 1 compared to 0.7 for antibiotic treated cultures. In E. paludosa, maximal cell abundance was higher in the presence of bacteria while the final carbon biomass did not vary, but in H. coffeaeformis maximal cell abundance did not vary significantly while final carbon biomass was higher in the presence of bacteria. By contrast, for both diatoms, cellular content of protein and lipids decrease significantly, as did extracellular carbon (EPS fraction) in the presence of bacteria. However, only a minor effect was observed on cellular carbohydrates, C/N ratio, and pigments (Chl a). Diatoms carbon fluxes towards the main biochemical components were also modified, with the protein carbon fraction significantly lower relative to other carbon compounds in the presence of high bacterial biomass. These results showed the complex interactions between diatoms and their associated bacteria. Promotion of diatoms growth by the presence of bacteria appears linked to change in microalgae biochemical composition that will modify the biofilm. Our results might help understanding the regulation of benthic biota in mudflat ecosystems

Heterotrophic Foraminifera Capable of Inorganic Nitrogen Assimilation

Nitrogen availability often limits biological productivity in marine systems, where inorganic nitrogen such as ammonium is assimilated into the food web by bacteria and photoautotrophic eukaryotes. Recently, ammonium assimilation was observed in kleptoplast-containing protists of the phylum foraminifera, possibly via the glutamine synthetase/glutamate synthase (GS/GOGAT) assimilation pathway imported with the kleptoplasts. However, it is not known if the ubiquitous and diverse heterotrophic protists have an innate ability for ammonium assimilation. Using stable isotope incubations (15N-ammonium and 13C-bicarbonate) and combining transmission electron microscopy (TEM) with quantitative nanoscale secondary ion mass spectrometry (NanoSIMS) imaging, we investigated the uptake and assimilation of dissolved inorganic ammonium by two heterotrophic foraminifera; a non-kleptoplastic benthic species, Ammonia sp., and a planktonic species, Globigerina bulloides. These species are heterotrophic and not capable of photosynthesis. Accordingly, they did not assimilate 13C-bicarbonate. However, both species assimilated dissolved 15N-ammonium and incorporated it into organelles of direct importance for ontogenetic growth and development of the cell. These observations demonstrate that at least some heterotrophic protists have an innate cellular mechanism for inorganic ammonium assimilation, highlighting a newly discovered pathway for dissolved inorganic nitrogen (DIN) assimilation within the marine microbial loop

Shorebirds affect ecosystem functioning on an intertidal mudflat

Ecosystem functioning and services have provided a rationale for conservation over the past decades. Intertidal muddy sediments, and the microphytobenthic biofilms that inhabit them, perform crucial ecosystem functions including erosion protection, nutrient cycling and carbon sequestration. It has been suggested that predation on sediment macrofauna by shorebirds may impact biofilms, and shorebirds are known to consume biofilm, potentially causing significant top-down effects on mudflat ecosystem functioning. We carried out an exclusion experiment on the Colne Estuary, Essex, UK to examine whether shorebird presence significantly affects sediment erodibility measured with a Cohesive Strength Meter (CSM) and microphytobenthos biomass measured using PAM fluorescence (Fo) and chlorophyll a content. We also tested for treatment effects on sediment-water nutrient fluxes (nitrate, nitrite, ammonia, phosphate and dissolved organic carbon (DOC)) during periods of both dark and light incubation. Excluding shorebirds caused statistically significant changes in regulating and provisioning ecosystem functions, including mudflat erodibility and nutrient fluxes. The presence of shorebirds lowered the sediment critical erosion threshold, reduced nitrate fluxes into the sediment under illumination, lowered nitrate efflux, and reduced phosphate uptake, compared to sediments where birds were excluded. There were no significant differences in macrofauna community composition within the sediment between treatments after 45 days of bird exclusion, suggesting a direct link between shorebird presence or absence and the significant differences in biofilm-related variables. This study introduces previously unknown effects of shorebird presence on ecosystem functions within this system and highlights an area of shorebird science that could aid joint conservation and human provisioning action

AZASPIRACIDS – Toxicological Evaluation, Test Methods and Identifcation of the Source Organisms (ASTOX II)

Since the Irish monitoring program was set up in 2001 azaspiracids (AZAs) have been detected in shellfish above the regulatory limit every year with the exception of 2004. The south west coast of Ireland is especially prone to the onsets of AZA events. Over this period a number of poisoning incidents associated with this toxin group have occurred, all related to Irish shellfish. In 2003 the Marine Institute was awarded funding for a research project named ASTOX. This project was very successful in producing a range of reference materials (RMs, which are essential for accurate detection and monitoring, and which up to this point were unavailable. The project also examined the toxicity of AZAs, primarily using in vitro cell assays but some in vivo studies were also performed. The overall aims of the ASTOX 2 project were to strengthen knowledge on the causative organism and toxicity of AZAs. The project aims were grouped into three areas: ecology, chemical support and toxicology.Marine Institute Marine Research Sub Programme (NDP 2007 - 2013), co financed under the European Regional Development Fund

Enrichment of intracellular sulphur cycle –associated bacteria in intertidal benthic foraminifera revealed by 16S and aprA gene analysis

Benthic foraminifera are known to play an important role in marine carbon and nitrogen cycles. Here, we report an enrichment of sulphur cycle -associated bacteria inside intertidal benthic foraminifera (Ammonia sp. (T6), Haynesina sp. (S16) and Elphidium sp. (S5)), using a meta barcoding approach targeting the 16S rRNA and aprA -genes. The most abundant intracellular bacterial groups included the genus Sulfurovum and the order Desulfobacterales. The bacterial 16S OTUs are likely to originate from the sediment bacterial communities, as the taxa found inside the foraminifera were also present in the sediment. The fact that 16S rRNA and aprA -gene derived intracellular bacterial OTUs were species-specific and significantly different from the ambient sediment community implies that bacterivory is an unlikely scenario, as benthic foraminifera are known to digest bacteria only randomly. Furthermore, these foraminiferal species are known to prefer other food sources than bacteria. The detection of sulphur-cycle related bacterial genes in this study suggests a putative role for these bacteria in the metabolism of the foraminiferal host. Future investigation into environmental conditions under which transcription of S-cycle genes are activated would enable assessment of their role and the potential foraminiferal/endobiont contribution to the sulphur-cycle.Peer reviewe