Micropearls and other intracellular inclusions of amorphous calcium carbonate: an unsuspected biomineralization capacity shared by diverse microorganisms.

An unsuspected biomineralization process, which produces intracellular inclusions of amorphous calcium carbonate (ACC), was recently discovered in unicellular eukaryotes. These mineral inclusions, called micropearls, can be highly enriched with other alkaline-earth metals (AEM) such as Sr and Ba. Similar intracellular inclusions of ACC have also been observed in prokaryotic organisms. These comparable biomineralization processes involving phylogenetically distant microorganisms are not entirely understood yet. This review gives a broad vision of the topic in order to establish a basis for discussion on the possible molecular processes behind the formation of the inclusions, their physiological role, the impact of these microorganisms on the geochemical cycles of AEM and their evolutionary relationship. Finally, some insights are provided to guide future research.

Morphological diversity and discrimination tools of the non-marine ostracod Cypridopsis silvestrii across temporal and spatial scales from Patagonia.

Geometric morphometric methods are powerful tools to discriminate between closely related ostracods taxa as well as to study the relationship between their morphological variations, taxonomy and paleoecology. In this study, valve outline analysis allows the discrimination between the non-marine ostracod C. silvestrii and R. whatleyi juveniles, pointing out differences in the posterior valve area and surface ornamentation. Modern female specimens of C. silvestrii from 23 sites located in a spatial transect (41 to 51 °S) exhibited extensive morphological variability, on the basis of which three morphotypes (acuminated, transitional, subtruncated) were determined. Multivariate analyses showed that acuminated and transitional shapes are not arranged in groups but the subtruncated morphotype, previously described as E. cecryphalium, seems to be associated with low water conductivity (372 µS cm-1), dominant cold (5 ºC) and windy (8.6 m s-1) climatic conditions. The fossil cluster which included nine cores spanning the last 15.6 kyr, only covered acuminated and transitional shapes, which may indicate that these lineages might be older than the subtruncated morphotype. In addition, morphological differences between reproduction modes suggested that parthenogenetic females exhibit de posterior margin more acuminate than sexual females. These results set the ground for more precise ecological and paleoenvironmental studies in Patagonia.

Characterizing ecoregions in Argentinian Patagonia using extant continental ostracods.

In order to characterize Patagonian (Argentina) ecoregions using non-marine ostracods, their associations in 69 environments were assessed. Twenty eight taxa were recorded, including 12 endemic of the Neotropical region. Our results indicate that Patagonian ostracods are mainly influenced by electrical conductivity (EC), altitude, pH, and temperature; and shows a correlation with Argentinian ecoregions. Assemblage I is composed of sites located at high altitude in the Andean Patagonian forest ecoregion. Host waters have low temperature, EC and pH, and support as representative species Cypris pubera, Eucypris virens, Bradleystrandesia fuscata, Tonacypris lutaria and Amphicypris nobilis. Assemblage II, related to mid-altitude environments in the Patagonian Steppe ecoregion, thrived in waters with moderate to high EC, and alkaline pH values. Dominant species includes Limnocythere rionegroensis, L. patagonica, E. virgata, Riocypris whatleyi, Riocypris sarsi, Newnhamia patagonica, Kapcypridopsis megapodus, Ilyocypris ramirezi and Penthesinelula incae. Assemblage III inhabited environments within Monte and Espinal ecoregions, situated in the eastern part of the study area at low altitude, EC moderate and temperate waters, supporting Heterocypris hyalinus, Amphicypris argentinensis, Sarscypridopsis aculeata, Cypridopsis vidua, Herpetocypris intermedia and Chlamidotheca incisa. Our results indicates that Argentinian Patagonia hosts a diverse ostracod fauna and highlights their capacity as proxies in ecological and palaeoenvironmental studies.

Strontium-90 pollution can be bioremediated with the green microalga Tetraselmis chui.

Strontium-90 (90Sr) is an artificial radioisotope produced by nuclear fission, with a relatively long half-life of 29 years. This radionuclide is released into the environment in the event of a nuclear incident, posing a serious risk to human and ecosystem health. There is a need to develop new efficient methods for the remediation of 90Sr, as current techniques for its removal have significant technical limitations and involve high energy and economic costs. Recently, several species of green microalgae within the class Chlorodendrophyceae have been found to form intracellular mineral inclusions of amorphous calcium carbonate (ACC), which can be highly enriched in natural (non-radiogenic) Sr. As bioremediation techniques are an attractive option to address radioactive pollution, we investigated the capacity of the unicellular alga Tetraselmis chui (class Chlorodendrophyceae) to sequester 90Sr. The 90Sr uptake capacity of T. chui cells was assessed in laboratory cultures by monitoring the time course of radioactivity in the culture medium using liquid scintillation counting (LSC). T. chui was shown to effectively sequester 90Sr, reducing the initial radioactivity of the culture medium by up to 50%. Thus, this study demonstrates the potential of the microalga T. chui to be used as a bioremediation agent against 90Sr pollution.

Prolonged drought periods over the last four decades increase flood intensity in southern Africa.

In semi-arid sub-Saharan Africa, climate change and the intensification of human activities have altered the hydrological balance and modified the recurrence of extreme hydroclimatic events, such as droughts and floods. The geomorphological heterogeneity of river catchments across the region, the variable human pressure, and the lack of continuous hydroclimatic data preclude the definition of proper mitigation strategies, with a direct effect on the sustainability of rural communities. Here, for the first time in Africa, we characterize hydrological extreme events using a multidisciplinary approach that includes sedimentary data from dams. We focus on the Limpopo River basin to evaluate which factors control flood magnitude since the 1970. Extreme flood events were identified across the basin in 1988-89, 1995-96, 1999-2000, 2003-04, 2010-11, 2013-14 and 2016-17. The statistical analysis of sedimentary flood records revealed a dramatic increase in their magnitude over the studied period. A positive correlation between maximum river flow and antecedent prolonged drought conditions was found in South Africa and Mozambique. Most importantly, since 1980, we observed the likely decoupling of extreme floods from the magnitude of La Niña events, suggesting that the natural interannual variability driven by El Niño-Southern Oscillation (ENSO) has been disrupted by climate changes and human activities.

High calcium and strontium uptake by the green microalga Tetraselmis chui is related to micropearl formation and cell growth.

Strontium-rich micropearls (intracellular inclusions of amorphous calcium carbonate) have been observed in several species of green microalgae within the class Chlorodendrophyceae, suggesting the potential use of these organisms for 90 Sr bioremediation purposes. However, very little is known about the micropearl formation process and the Ca and Sr uptake dynamics of these microalgae. To better understand this phenomenon, we investigated, through laboratory cultures, the behaviour of two species within the class Chorodendrophyceae: Tetraselmis chui, forming micropearls, and T. marina, not forming micropearls. We show that T. chui growth and micropearl formation requires available Ca in the culture medium, and that the addition of dissolved Sr can partially replace the function of Ca in cells. On the other hand, T. marina can grow without added Ca and Sr, probably due to its inability to form micropearls. T. chui cells show a high Ca and Sr uptake, significantly decreasing the concentration of both elements in the culture medium. Strontium is incorporated in micropearls in a short period of time, suggesting that micropearl formation is, most likely, a fast process that only takes a few hours. In addition, we show that micropearls equally distribute between daughter cells during cell division.

Hydrochemistry, isotope studies and salt formation in saline lakes of arid regions: Extra-Andean Patagonia, Argentina.

The most favourable locations for the development of saline lakes are in the rain-shadow of mountain ranges, which provide large areas of precipitation catchment while the base of the basin is under arid climate and exposed to evaporation. These conditions are found in Extra-Andean Patagonia under the rain-shadow generated by the Andean cordillera. There, an endorheic basin with two shallow and saline lakes, Cari Laufquen Chica (CLC) and Cari Laufquen Grande (CLG), was studied with the aim of analysing the factors that condition the hydrochemical processes acting in the formation of evaporites associated with these environments. A monitoring network was installed and five surface and groundwater survey campaigns were carried out at different points in the basin to define groundwater flow and also to extract samples. In situ pH, electrical conductivity and temperature and laboratory physicochemical determinations of major ions and stable isotopes of the water were measured. SEM-EDS and XRD analyses of saline crusts from the edge of the lakes and adjacent sediments were carried out. The obtained results allowed identifying that CLC and CLG saline lakes have different surface water-groundwater configurations. CLC is a flow-through shallow lake, while CLG is a discharge shallow lake. The analysis of the saline precipitates and the chemical facies of the water allows identifying cycles of dissolution, evaporation and precipitation in both saline lakes. However, the different groundwater flow between the two lakes is reflected in their salinity as well as in the evolution and development of saline precipitates. CLC saline lake shows a dissolution-precipitation trend of thenardite and Ca-Mg carbonates, whereas CLG saline lake displays a trend of dissolution-precipitation of halite, thenardite and trona with strong dominance of halite. The present study identifies for the first time the main factors conditioning hydrochemical processes in these saline lakes of extra-Andean Patagonia. Our results indicate that the hydrological configuration with respect to the groundwater flow is the prevailing factor setting the hydrochemical processes that trigger the formation of salt crusts.

Intensified microbial sulfate reduction in the deep Dead Sea during the early Holocene Mediterranean sapropel 1 deposition.

The hypersaline Dead Sea and its sediments are natural laboratories for studying extremophile microorganism habitat response to environmental change. In modern times, increased freshwater runoff to the lake surface waters resulted in stratification and dilution of the upper water column followed by microbial blooms. However, whether these events facilitated a microbial response in the deep lake and sediments is obscure. Here we investigate archived evidence of microbial processes and changing regional hydroclimate conditions by reconstructing deep Dead Sea chemical compositions from pore fluid major ion concentration and stable S, O, and C isotopes, together with lipid biomarkers preserved in the hypersaline deep Dead Sea ICDP-drilled core sediments dating to the early Holocene (ca. 10,000 years BP). Following a significant negative lake water balance resulting in salt layer deposits at the start of the Holocene, there was a general period of positive net water balance at 9500-8300 years BP. The pore fluid isotopic composition of sulfate exhibit evidence of intensified microbial sulfate reduction, where both δ34S and δ18O of sulfate show a sharp increase from estimated base values of 15.0‰ and 13.9‰ to 40.2‰ and 20.4‰, respectively, and a δ34S vs. δ18O slope of 0.26. The presence of the n-C17 alkane biomarker in the sediments suggests an increase of cyanobacteria or phytoplankton contribution to the bulk organic matter that reached the deepest parts of the Dead Sea. Although hydrologically disconnected, both the Mediterranean Sea and the Dead Sea microbial ecosystems responded to increased freshwater runoff during the early Holocene, with the former depositing the organic-rich sapropel 1 layer due to anoxic water column conditions. In the Dead Sea prolonged positive net water balance facilitated primary production and algal blooms in the upper waters and intensified microbial sulfate reduction in the hypolimnion and/or at the sediment-brine interface.

Organic matter mineralization in modern and ancient ferruginous sediments.

Deposition of ferruginous sediment was widespread during the Archaean and Proterozoic Eons, playing an important role in global biogeochemical cycling. Knowledge of organic matter mineralization in such sediment, however, remains mostly conceptual, as modern ferruginous analogs are largely unstudied. Here we show that in sediment of ferruginous Lake Towuti, Indonesia, methanogenesis dominates organic matter mineralization despite highly abundant reactive ferric iron phases like goethite that persist throughout the sediment. Ferric iron can thus be buried over geologic timescales even in the presence of labile organic carbon. Coexistence of ferric iron with millimolar concentrations of methane further demonstrates lack of iron-dependent methane oxidation. With negligible methane oxidation, methane diffuses from the sediment into overlying waters where it can be oxidized with oxygen or escape to the atmosphere. In low-oxygen ferruginous Archaean and Proterozoic oceans, therefore, sedimentary methane production was likely favored with strong potential to influence Earth's early climate.

Weak Influence of Paleoenvironmental Conditions on the Subsurface Biosphere of Lake Ohrid over the Last 515 ka.

Lacustrine sediments are widely used to investigate the impact of climatic change on biogeochemical cycling. In these sediments, subsurface microbial communities are major actors of this cycling but can also affect the sedimentary record and overprint the original paleoenvironmental signal. We therefore investigated the subsurface microbial communities of the oldest lake in Europe, Lake Ohrid (North Macedonia, Albania), to assess the potential connection between microbial diversity and past environmental change using 16S rRNA gene sequences. Along the upper ca. 200 m of the DEEP site sediment record spanning ca. 515 thousand years (ka), our results show that Atribacteria, Bathyarchaeia and Gammaproteobacteria structured the community independently from each other. Except for the latter, these taxa are common in deep lacustrine and marine sediments due to their metabolic versatility adapted to low energy environments. Gammaproteobacteria were often co-occurring with cyanobacterial sequences or soil-related OTUs suggesting preservation of ancient DNA from the water column or catchment back to at least 340 ka, particularly in dry glacial intervals. We found significant environmental parameters influencing the overall microbial community distribution, but no strong relationship with given phylotypes and paleoclimatic signals or sediment age. Our results support a weak recording of early diagenetic processes and their actors by bulk prokaryotic sedimentary DNA in Lake Ohrid, replaced by specialized low-energy clades of the deep biosphere and a marked imprint of erosional processes on the subsurface DNA pool of Lake Ohrid.

Planktonic foraminifera eDNA signature deposited on the seafloor remains preserved after burial in marine sediments.

Environmental DNA (eDNA) metabarcoding of marine sediments has revealed large amounts of sequences assigned to planktonic taxa. How this planktonic eDNA is delivered on the seafloor and preserved in the sediment is not well understood. We address these questions by comparing metabarcoding and microfossil foraminifera assemblages in sediment cores taken off Newfoundland across a strong ecological gradient. We detected planktonic foraminifera eDNA down to 30 cm and observed that the planktonic/benthic amplicon ratio changed with depth. The relative proportion of planktonic foraminiferal amplicons remained low from the surface down to 10 cm, likely due to the presence of DNA from living benthic foraminifera. Below 10 cm, the relative proportion of planktonic foraminifera amplicons rocketed, likely reflecting the higher proportion of planktonic eDNA in the DNA burial flux. In addition, the microfossil and metabarcoding assemblages showed a congruent pattern indicating that planktonic foraminifera eDNA is deposited without substantial lateral advection and preserves regional biogeographical patterns, indicating deposition by a similar mechanism as the foraminiferal shells. Our study shows that the planktonic eDNA preserved in marine sediments has the potential to record climatic and biotic changes in the pelagic community with the same spatial and temporal resolution as microfossils.

Biomineralization Capacities of Chlorodendrophyceae: Correlation Between Chloroplast Morphology and the Distribution of Micropearls in the Cell.

Several species of the genus Tetraselmis (Chlorodendrophyceae, Chlorophyta) were recently discovered to possess unsuspected biomineralization capacities: they produce multiple intracellular inclusions of amorphous calcium carbonate (ACC), called micropearls. Early light-microscopists had spotted rows of refractive granules in some species, although without identifying their mineral nature. Scanning electron microscope (SEM) observations showed that the distribution of the micropearls in the cell forms a pattern, which appears to be characteristic for a given species. The present study shows that this pattern correlates with the shape of the chloroplast, which differs between Tetraselmis species, because micropearls align themselves along the incisions between chloroplast lobes. This was observed both by SEM and in live cells by light microscopy (LM) using Nomarski differential interference contrast. Additionally, molecular phylogenetic analyses, of rbcL and ITS2 gene sequences from diverse strains of Chlorodendrophyceae, corroborated the morphological observations by identifying two groups among nominal Tetraselmis spp. that differ in chloroplast morphology, micropearl arrangement, and ITS2 RNA secondary structure.

Microbial community composition along a 50 000-year lacustrine sediment sequence.

For decades, microbial community composition in subseafloor sediments has been the focus of extensive studies. In deep lacustrine sediments, however, the taxonomic composition of microbial communities remains undercharacterized. Greater knowledge on microbial diversity in lacustrine sediments would improve our understanding of how environmental factors, and resulting selective pressures, shape subsurface biospheres in marine and freshwater sediments. Using high-throughput sequencing of 16S rRNA genes across high-resolution climate intervals covering the last 50 000 years in Laguna Potrok Aike, Argentina, we identified changes in microbial populations in response to both past environmental conditions and geochemical changes of the sediment during burial. Microbial communities in Holocene sediments were most diverse, reflecting a layering of taxa linked to electron acceptors availability. In deeper intervals, the data show that salinity, organic matter and the depositional conditions over the Last Glacial-interglacial cycle were all selective pressures in the deep lacustrine assemblage resulting in a genetically distinct biosphere from the surface dominated primarily by Bathyarchaeota and Atribacteria groups. However, similar to marine sediments, some dominant taxa in the shallow subsurface persisted into the subsurface as minor fraction of the community. The subsequent establishment of a deep subsurface community likely results from a combination of paleoenvironmental factors that have shaped the pool of available substrates, together with substrate depletion and/or reworking of organic matter with depth.