Rapid Freezing and Cryo-SEM-EDS Imaging of Foraminifera (Unicellular Eukaryotes) Using a Conductive Viscous Cryogenic Glue.

Spatial distribution of water-soluble molecules and ions in living organisms is still challenging to assess. Energy-dispersive X-ray spectroscopy (EDS) via cryogenic scanning electron microscopy (cryo-SEM) is one of the promising methods to study them without loss of dissolved contents. High-resolution cryo-SEM-EDS has challenges in sample preparation, including cross-section exposure and sample drift/charging due to insulative surrounding water. The former becomes problematic for large and inseparable organisms, such as benthic foraminifera, a unicellular eukaryote playing significant roles in marine ecosystems, which often exceed the size limit for the most reliable high-pressure freezing. Here we show graphite oxide dispersed in sucrose solution as a good glue to freeze, expose cross-section by cryo-ultramicrotome, and analyze elemental distribution owing to the glue's high viscosity, adhesion force, and electron conductivity. To demonstrate the effectiveness and applicability of the glue for cryo-SEM-EDS, deep-sea foraminifer Uvigerina akitaensis was sampled during a cruise and plunge frozen directly on the research vessel, where the liquid nitrogen supply is limited. The microstructures were preserved as faithfully in cryo-SEM images as those with the conventional resin-substituted transmission electron micrograph. We found elements colocalized within the cytoplasm originating from water-soluble compounds that can be lost with conventional dehydrative fixation.

Fast and pervasive diagenetic isotope exchange in foraminifera tests is species-dependent.

Oxygen isotope compositions of fossil foraminifera tests are commonly used proxies for ocean paleotemperatures, with reconstructions spanning the last 112 million years. However, the isotopic composition of these calcitic tests can be substantially altered during diagenesis without discernible textural changes. Here, we investigate fluid-mediated isotopic exchange in pristine tests of three modern benthic foraminifera species (Ammonia sp., Haynesina germanica, and Amphistegina lessonii) following immersion into an 18O-enriched artificial seawater at 90 °C for hours to days. Reacted tests remain texturally pristine but their bulk oxygen isotope compositions reveal rapid and species-dependent isotopic exchange with the water. NanoSIMS imaging reveals the 3-dimensional intra-test distributions of 18O-enrichment that correlates with test ultra-structure and associated organic matter. Image analysis is used to quantify species level differences in test ultrastructure, which explains the observed species-dependent rates of isotopic exchange. Consequently, even tests considered texturally pristine for paleo-climatic reconstruction purposes may have experienced substantial isotopic exchange; critical paleo-temperature record re-examination is warranted.

Test structure in some pioneer multichambered Paleozoic foraminifera.

Foraminiferal wall microstructures, consistent with the molecular-based high-rank classification, are critical to understanding foraminiferal evolution and advanced taxonomic relationships. Although test structures are well documented for recent, Cenozoic, and some Mesozoic foraminifera, the diagnostic characteristics of Paleozoic taxa are largely unexplored. The majority of calcareous Paleozoic foraminifera have been assigned to the Fusulinata based on questionable homogeneously "microgranular" test wall microstructures, which have never been sufficiently documented for most taxa. We investigated the test structures of exceptionally well-preserved Devonian (Eifelian) Semitextularia thomasi, representing the first calcareous true multichambered (serial) foraminifera, and compared this species with a large fusiform Permian representative of "true" fusulinids (Neoschwagerinidae). The tests of Semitextularia thomasi display lamellar structures that are not observed in any other fossil or recent foraminiferal group. The Paleozoic foraminifera, traditionally referred to one taxon (the class Fusulinata), possess at least three contrasting test wall microstructures, representing separate high-rank taxonomic groups. Fusulinata is most likely a highly polyphyletic group that is in need of taxonomic revision. The term Fusulinata, defined as including all Paleozoic calcareous forms except Miliolida and Lagenata, is not phylogenetically meaningful and should no longer be used or should be restricted to true complex fusulinids with microgranular test structures, which appeared in the Carboniferous.

Calcite crystal orientation patterns in the bilayers of laminated shells of benthic rotaliid foraminifera.

Shells of calcifying foraminifera play a major role in marine biogeochemical cycles; fossil shells form important archives for paleoenvironment reconstruction. Despite their importance in many Earth science disciplines, there is still little consensus on foraminiferal shell mineralization. Geochemical, biochemical, and physiological studies showed that foraminiferal shell formation might take place through various and diverse mineralization mechanisms. In this study, we contribute to benthic foraminiferal shell calcification through deciphering crystallite organization within the shells. We base our conclusions on results gained from electron backscattered diffraction (EBSD) measurements and describe microstructure/texture characteristics within the laminated shell walls of the benthic, symbiontic foraminifera: Ammonia tepida, Amphistegina lobifera, Amphistegina lessonii. We highlight crystallite assembly patterns obtained on differently oriented cuts and discuss crystallite sizes, morphologies, interlinkages, orientations, and co-orientation strengths. We show that: (i) crystals within benthic foraminiferal shells are mesocrystals, (ii) have dendritic-fractal morphologies and (iii) interdigitate strongly. Based on crystal size, we (iv) differentiate between the two layers that comprise the shells and demonstrate that (v) crystals in the septa have different assemblies relative to those in the shell walls. We highlight that (vi) at junctions of different shell elements the axis of crystal orientation jumps abruptly such that their assembly in EBSD maps has a bimodal distribution. We prove (vii) extensive twin-formation within foraminiferal calcite; we demonstrate (viii) the presence of two twin modes: 60°/[001] and 77°/~[6 -6 1] and visualize their distributions within the shells. In a broader perspective, we draw conclusions on processes that lead to the observed microstructure/texture patterns.

Integrating morphology and metagenomics to understand taxonomic variability of Amphisorus (Foraminifera, Miliolida) from Western Australia and Indonesia.

Foraminifera are a group of mostly marine protists with high taxonomic diversity. Species identification is often complex, as both morphological and molecular approaches can be challenging due to a lack of unique characters and reference sequences. An integrative approach combining state of the art morphological and molecular tools is therefore promising. In this study, we analysed large benthic Foraminifera of the genus Amphisorus from Western Australia and Indonesia. Based on previous findings on high morphological variability observed in the Soritidae and the discontinuous distribution of Amphisorus along the coast of western Australia, we expected to find multiple morphologically and genetically unique Amphisorus types. In order to gain detailed insights into the diversity of Amphisorus, we applied micro CT scanning and shotgun metagenomic sequencing. We identified four distinct morphotypes of Amphisorus, two each in Australia and Indonesia, and showed that each morphotype is a distinct genotype. Furthermore, metagenomics revealed the presence of three dinoflagellate symbiont clades. The most common symbiont was Fugacium Fr5, and we could show that its genotypes were mostly specific to Amphisorus morphotypes. Finally, we assembled the microbial taxa associated with the two Western Australian morphotypes, and analysed their microbial community composition. Even though each Amphisorus morphotype harboured distinct bacterial communities, sampling location had a stronger influence on bacterial community composition, and we infer that the prokaryotic community is primarily shaped by the microhabitat rather than host identity. The integrated approach combining analyses of host morphology and genetics, dinoflagellate symbionts, and associated microbes leads to the conclusion that we identified distinct, yet undescribed taxa of Amphisorus. We argue that the combination of morphological and molecular methods provides unprecedented insights into the diversity of foraminifera, which paves the way for a deeper understanding of their biodiversity, and facilitates future taxonomic and ecological work.

Abundant Chitinous Structures in Chilostomella (Foraminifera, Rhizaria) and Their Potential Functions.

Benthic foraminifera, members of Rhizaria, inhabit a broad range of marine environments and are particularly common in hypoxic sediments. The biology of benthic foraminifera is key to understanding benthic ecosystems and relevant biogeochemical cycles, especially in hypoxic environments. Chilostomella is a foraminiferal genus commonly found in hypoxic deep-sea sediments and has poorly understood ecological characteristics. For example, the carbon isotopic compositions of their lipids are substantially different from other co-occurring genera, probably reflecting unique features of its metabolism. Here, we investigated the cytoplasmic and ultrastructural features of Chilostomella ovoidea from bathyal sediments of Sagami Bay, Japan, based on serial semi-thin sections examined using an optical microscope followed by a three-dimensional reconstruction, combined with TEM observations of ultra-thin sections. Observations by TEM revealed the presence of abundant electron-dense structures dividing the cytoplasm. Based on histochemical staining, these structures are shown to be composed of chitin. Our 3D reconstruction revealed chitinous structures in the final seven chambers. These exhibited a plate-like morphology in the final chambers but became rolled up in earlier chambers (toward the proloculus). These chitinous, plate-like structures may function to partition the cytoplasm in a chamber to increase the surface/volume ratio and/or act as a reactive site for some metabolic functions.

Nanoscale trace metal imprinting of biocalcification of planktic foraminifers by Toba's super-eruption.

Bioactive metal releases in ocean surface water, such as those by ash falls during volcanic super-eruptions, might have a potentially toxic impact on biocalcifier planktic microorganisms. Nano-XRF imaging with the cutting-edge synchrotron hard X-ray nano-analysis ID16B beamline (ESRF) revealed for the first time a specific Zn- and Mn-rich banding pattern in the test walls of Globorotalia menardii planktic foraminifers extracted from the Young Toba Tuff layer, and thus contemporaneous with Toba's super-eruption, 74,000 years ago. The intra-test correlation of Zn and Mn patterns at the nanoscale with the layered calcareous microarchitecture, indicates that the incorporation of these metals is syngenetic to the wall growth. The preferential Mn and Zn sequestration within the incipient stages of chamber formation suggests a selective incorporation mechanism providing a resilience strategy to metal pollution in the test building of planktic foraminifers.

Unassigned diversity of planktonic foraminifera from environmental sequencing revealed as known but neglected species.

Most research on extant planktonic foraminifera has been directed towards larger species (>0.150 mm) which can be easily manipulated, counted and yield enough calcite for geochemical analyses. This has drawn attention towards the macroperforate clade and created an impression of their numerical and ecological dominance. Drawing such conclusions from the study of such "giants" is a dangerous path. There were times in the evolutionary history of planktonic foraminifera when all species were smaller than 0.1 mm and indeed numerous small taxa, mainly from the microperforate clade, have been formally described from the modern plankton. The significance of these small, obscure and neglected species is poorly characterized and their relationship to the newly discovered hyperabundant but uncharacterized lineages of planktonic foraminifera in metabarcoding datasets is unknown. To determine, who is hiding in the metabarcoding datasets, we carried out an extensive sequencing of 18S rDNA targeted at small and obscure species. The sequences of the newly characterized small and obscure taxa match many of the previously uncharacterized lineages found in metabarcoding data. This indicates that most of the modern diversity in planktonic foraminifera has been taxonomically captured, but the role of the small and neglected taxa has been severely underestimated.

Morphological diversity in the foraminiferal genus Marginopora.

Benthic foraminifera, and certainly symbiont-bearing (large) benthic foraminifera are generally considered to have large geographic ranges in combination with significant ecomorphological variation. With the advance of molecular phylogenetic approaches, supported or preceded by detailed morphological studies, it was demonstrated that this view needs to be reevaluated. In this paper I evaluate the morphology of five Marginopora populations from around the Coral Sea by microCT-scanning. I argue that ecomorphological and ontogenetic variation is smaller than geographic variation in morphology. This forms the basis for the description of three new species, M. santoensis nov. spec., M. charlottensis nov. spec., M. orpheusensis nov. spec. Quantitative morphological variation between M. rossi, M. orpheusensis nov. spec. and M. charlottensis nov. spec. is overlapping, but each species has unique morphological characters supporting recognition as new species. Support to distinguish the deep living (M. rossi, M. charlottensis nov. spec., M. orpheusensis nov. spec.) and shallow living (M. vertebralis) Marginopora populations as separate species is strong, but not enough molecular phylogenetic data are available to test the three new deep-living species on the Great Barrier Reef hypothesis. However, detailed understanding of ecophenotypic variation in M. santoensis nov. spec. supports the conclusion that it is unlikely that ecophenotypic variation can explain the morphological variation between the three species. I argue that the number of species in this genus is underestimated, and that there are at least five species in the Coral Sea area alone.

Micro-CT 3D imaging reveals the internal structure of three abyssal xenophyophore species (Protista, Foraminifera) from the eastern equatorial Pacific Ocean.

Xenophyophores, giant foraminifera, are distinctive members of the deep-sea megafauna that accumulate large masses of waste material ('stercomare') within their agglutinated tests, and organise their cells as branching strands enclosed within an organic tube (the 'granellare' system). Using non-destructive, three-dimensional micro-CT imaging we explored these structures in three species from the abyssal eastern Pacific Clarion-Clipperton Zone (CCZ). In Psammina spp., the low-density stercomare occupied much of the test interior, while high-density granellare strands branched throughout the structure. In Galatheammina sp. the test comprised a mixture of stercomare and test particles, with the granellare forming a web-like system of filaments. The granellare occupied 2.8-5.1%, the stercomare 72.4-82.4%, and test particles 14.7-22.5%, of the 'body' volume in the two Psammina species. The corresponding proportions in Galatheammina sp. were 1.7% (granellare), 39.5% (stercomare) and 58.8% (test particles). These data provide a potential basis for estimating the contribution of xenophyophores to seafloor biomass in areas like the CCZ where they dominate the megafauna. As in most xenophyophore species, the granellare hosted huge numbers of tiny barite crystals. We speculate that these help to support the extensive granellare system, as well as reducing the cell volume and lightening the metabolic burden required to maintain it.

Keystone Arctic paleoceanographic proxy association with putative methanotrophic bacteria.

Foraminifera in sediments exposed to gas-hydrate dissociation are not expected to have cellular adaptations that facilitate inhabitation of chemosynthesis-based ecosystems because, to date, there are no known endemic seep foraminifera. To establish if foraminifera inhabit sediments impacted by gas-hydrate dissociation, we examined the cellular ultrastructure of Melonis barleeanus (Williamson, 1858) from the Vestnesa gas hydrate province (Arctic Ocean, west of Svalbard at ~79 °N; ~1200-m depth; n = 4). From sediments with gas hydrate indicators, living M. barleeanus had unusual pore plugs composed of a thick, fibrous meshwork; mitochondria were concentrated at the cell periphery, under pore plugs. While there was no evidence of endosymbioses with prokaryotes, most M. barleeanus specimens were associated with what appear to be Type I methanotrophic bacteria. One foraminifer had a particularly large bolus of these microbes concentrated near its aperture. This is the first documented instance of bona fide living M. barleeanus in gas-hydrate sediments and first documentation of a foraminifer living in close association with putative methanotrophs. Our observations have implications to paleoclimate records utilizing this foundational foraminiferal species.

Unlocking the biomineralization style and affinity of Paleozoic fusulinid foraminifera.

Fusulinids are the most diverse, abundant and geographically widespread Paleozoic foraminifera which are widely considered to possess a "homogeneously microgranular" test microstructure composed of subangular grains of several micrometers in size. However, this texture appears to be a diagenetic artifact. Here we describe well-preserved Devonian calcareous fusulinids (Nanicella) from the Holy Cross Mountains (HCM) in central Poland. Foraminifera from Poland in which the primary nature of tests have not been masked by diagenesis are composed of low magnesium calcite spherical grains up to about 100 nanometers in diameter, identical to those observed in Recent and fossil hyaline foraminifera (Rotaliida, Globothalamea). These data call the paradigm of microgranular test microstructure of Foraminifera into question, and suggest a possible phylogenetic relationship between globothalamids and some fusulinids.

Ostracoda and foraminifera from Paleocene (Olinda well), Paraíba Basin, Brazilian Northeast

ABSTRACT Paleocene ostracods and planktonic foraminifera from the Maria Farinha Formation, Paraíba Basin, are herein presented. Eleven ostracod species were identified in the genera Cytherella Jones, Cytherelloidea Alexander, Eocytheropteron Alexander, Semicytherura Wagner, Paracosta Siddiqui, Buntonia Howe, Soudanella Apostolescu, Leguminocythereis Howe and, probably, Pataviella Liebau. The planktonic foraminifera are represented by the genera Guembelitria Cushman, Parvularugoglobigerina Hofker, Woodringina Loeblich and Tappan, Heterohelix Ehrenberg, Zeauvigerina Finlay, Muricohedbergella Huber and Leckie, and Praemurica Olsson, Hemleben, Berggren and Liu. The ostracods and foraminifera analyzed indicate an inner shelf paleoenvironment for the studied section. Blooms of Guembelitria spp., which indicate either shallow environments or upwelling zones, were also recorded reinforcing previous paleoenvironmental interpretations based on other fossil groups for this basin.

Ostracoda and foraminifera from Paleocene (Olinda well), Paraíba Basin, Brazilian Northeast.

Paleocene ostracods and planktonic foraminifera from the Maria Farinha Formation, Paraíba Basin, are herein presented. Eleven ostracod species were identified in the genera Cytherella Jones, Cytherelloidea Alexander, Eocytheropteron Alexander, Semicytherura Wagner, Paracosta Siddiqui, Buntonia Howe, Soudanella Apostolescu, Leguminocythereis Howe and, probably, Pataviella Liebau. The planktonic foraminifera are represented by the genera Guembelitria Cushman, Parvularugoglobigerina Hofker, Woodringina Loeblich and Tappan, Heterohelix Ehrenberg, Zeauvigerina Finlay, Muricohedbergella Huber and Leckie, and Praemurica Olsson, Hemleben, Berggren and Liu. The ostracods and foraminifera analyzed indicate an inner shelf paleoenvironment for the studied section. Blooms of Guembelitria spp., which indicate either shallow environments or upwelling zones, were also recorded reinforcing previous paleoenvironmental interpretations based on other fossil groups for this basin.

Benthic Foraminifera from the Capricorn Group, Great Barrier Reef, Australia.

Effective reef management and monitoring has become increasingly important as anthropogenic processes impact upon natural ecosystems. One locality that is under direct threat due to human activities is the Australian Great Barrier Reef (GBR). Marine foraminifera represent an abundant and readily applicable tool that can be used in reef studies to investigate a variety of ecological parameters and assist in understanding reef dynamics and influence management protocols. The first step is to establish a baseline knowledge of taxonomic composition within the region to facilitate comparative studies and monitor how assemblages change in order to maximise effective management. A detailed taxonomic assessment is provided of 133 species of benthic foraminifera in 76 genera from Heron Island, One Tree Island, Wistari and Sykes Reefs, which form the core of the Capricorn Group (CG) at the southern end of the GBR. Of these 133 species, 46% belong to the order Miliolida, 34% to Rotaliida, 7% to Textulariida, 5% to Lagenida, 3% to Lituolida, 3% to Spirillinida, 1% to Loftusiida and 1% to Robertinida. Samples were collected from a variety of shallow shelf reef environments including reef flat, lagoonal and channel environments. Seventy species, representing the most abundant forms, are formally described with detailed distribution data for the remaining 63 species supplied.

Biomineralization pathways in a foraminifer revealed using a novel correlative cryo-fluorescence-SEM-EDS technique.

Foraminifera are marine protozoans that are widespread in oceans throughout the world. Understanding biomineralization pathways in foraminifera is particularly important because their calcitic shells are major components of global calcium carbonate production. We introduce here a novel correlative approach combining cryo-SEM, cryo-fluorescence imaging and cryo-EDS. This approach is applied to the study of ion transport processes in the benthic foraminifer genus Amphistegina. We confirm the presence of large sea water vacuoles previously identified in intact and partially decalcified Amphistegina lobifera specimens. We observed relatively small vesicles that were labelled strongly with calcein, and also identified magnesium (Mg)-rich mineral particles in the cytoplasm, as well as in the large sea water vacuoles. The combination of cryo-microscopy with elemental microanalysis and fluorescence imaging reveals new aspects of the biomineralization pathway in foraminifera which are, to date, unique in the world of biomineralization. This approach is equally applicable to the study of biomineralization pathways in other organisms.

Spatial Patterns in the Distribution, Diversity and Abundance of Benthic Foraminifera around Moorea (Society Archipelago, French Polynesia).

Coral reefs are now subject to global threats and influences from numerous anthropogenic sources. Foraminifera, a group of unicellular shelled organisms, are excellent indicators of water quality and reef health. Thus we studied a set of samples taken in 1992 to provide a foraminiferal baseline for future studies of environmental change. Our study provides the first island-wide analysis of shallow benthic foraminifera from around Moorea (Society Archipelago). We analyzed the composition, species richness, patterns of distribution and abundance of unstained foraminiferal assemblages from bays, fringing reefs, nearshore and back- and fore-reef environments. A total of 380 taxa of foraminifera were recorded, a number that almost doubles previous species counts. Spatial patterns of foraminiferal assemblages are characterized by numerical abundances of individual taxa, cluster groups and gradients of species richness, as documented by cluster, Fisher α, ternary plot and Principal Component Analyses (PCA). The inner bay inlets are dominated by stress-tolerant, mostly thin-shelled taxa of Bolivina, Bolivinella, Nonionoides, Elongobula, and Ammonia preferring low-oxygen and/or nutrient-rich habitats influenced by coastal factors such as fresh-water runoff and overhanging mangroves. The larger symbiont-bearing foraminifera (Borelis, Amphistegina, Heterostegina, Peneroplis) generally live in the oligotrophic, well-lit back- and fore-reef environments. Amphisteginids and peneroplids were among the few taxa found in the bay environments, probably due to their preferences for phytal substrates and tolerance to moderate levels of eutrophication. The fringing reef environments along the outer bay are characterized by Borelis schlumbergeri, Heterostegina depressa, Textularia spp. and various miliolids which represent a hotspot of diversity within the complex reef-lagoon system of Moorea. The high foraminiferal Fisher α and species richness diversity in outer bay fringing reefs is consistent with the disturbance-mosaic (microhabitat heterogeneity) hypothesis. Calculations of the FORAM Index (FI), a single metric index to assess reef vitality, indicate that all fore- and most back-reef environments support active carbonate accretion and provide habitat suitability for carbonate producers dependent on algal symbiosis. Lowest suitability values were recorded within the innermost bays, an area where natural and increasing anthropogenic influences continue to impact the reefs. The presence of habitat specific assemblages and numerical abundance values of individual taxa show that benthic foraminifera are excellent recorders of environmental perturbations and good indicators useful in modern and ancient ecological and environmental studies.

Fossil and genetic evidence for the polyphyletic nature of the planktonic foraminifera "Globigerinoides", and description of the new genus Trilobatus.

Planktonic foraminifera are one of the most abundant and diverse protists in the oceans. Their utility as paleo proxies requires rigorous taxonomy and comparison with living and genetically related counterparts. We merge genetic and fossil evidence of "Globigerinoides", characterized by supplementary apertures on spiral side, in a new approach to trace their "total evidence phylogeny" since their first appearance in the latest Paleogene. Combined fossil and molecular genetic data indicate that this genus, as traditionally understood, is polyphyletic. Both datasets indicate the existence of two distinct lineages that evolved independently. One group includes "Globigerinoides" trilobus and its descendants, the extant "Globigerinoides" sacculifer, Orbulina universa and Sphaeroidinella dehiscens. The second group includes the Globigerinoides ruber clade with the extant G. conglobatus and G. elongatus and ancestors. In molecular phylogenies, the trilobus group is not the sister taxon of the ruber group. The ruber group clusters consistently together with the modern Globoturborotalita rubescens as a sister taxon. The re-analysis of the fossil record indicates that the first "Globigerinoides" in the late Oligocene are ancestral to the trilobus group, whereas the ruber group first appeared at the base of the Miocene with representatives distinct from the trilobus group. Therefore, polyphyly of the genus "Globigerinoides" as currently defined can only be avoided either by broadening the genus concept to include G. rubescens and a large number of fossil species without supplementary apertures, or if the trilobus group is assigned to a separate genus. Since the former is not feasible due to the lack of a clear diagnosis for such a broad genus, we erect a new genus Trilobatus for the trilobus group (type species Globigerina triloba Reuss) and amend Globoturborotalita and Globigerinoides to clarify morphology and wall textures of these genera. In the new concept, Trilobatus n. gen. is paraphyletic and gave rise to the Praeorbulina/Orbulina and Sphaeroidinellopsis/Sphaeroidinella lineages.

Effects of lead pollution on Ammonia parkinsoniana (foraminifera): ultrastructural and microanalytical approaches.

The responses of Ammonia parkinsoniana (Foraminifera) exposed to different concentrations of lead (Pb) were evaluated at the cytological level. Foraminifera-bearing sediments were placed in mesocosms that were housed in aquaria each with seawater of a different lead concentration. On the basis of transmission electron microscopy and environmental scanning electron microscopy coupled with energy dispersive spectrometer analyses, it was possible to recognize numerous morphological differences between untreated (i.e., control) and treated (i.e., lead enrichment) specimens. In particular, higher concentrations of this pollutant led to numerical increase of lipid droplets characterized by a more electron-dense core, proliferation of residual bodies, a thickening of the organic lining, mitochondrial degeneration, autophagosome proliferation and the development of inorganic aggregates.  All these cytological modifications might be related to the pollutant-induced stress and some of them such as the thickening of organic lining might suggest a potential mechanism of protection adopted by foraminifera.

The impact of ocean acidification on the functional morphology of foraminifera.

Culturing experiments were performed on sediment samples from the Ythan Estuary, N. E. Scotland, to assess the impacts of ocean acidification on test surface ornamentation in the benthic foraminifer Haynesina germanica. Specimens were cultured for 36 weeks at either 380, 750 or 1000 ppm atmospheric CO2. Analysis of the test surface using SEM imaging reveals sensitivity of functionally important ornamentation associated with feeding to changing seawater CO2 levels. Specimens incubated at high CO2 levels displayed evidence of shell dissolution, a significant reduction and deformation of ornamentation. It is clear that these calcifying organisms are likely to be vulnerable to ocean acidification. A reduction in functionally important ornamentation could lead to a reduction in feeding efficiency with consequent impacts on this organism's survival and fitness.