Fossil amoebae (Hemiarcherellidae fam. nov.) from Albian (Cretaceous) amber of France

Abstract: Two extraordinarily well‐preserved testate amoebae are described from Late Albian age amber from south‐western France. The specimens are attributed to a new family, the Hemiarcherellidae fam. nov., and are described as Hemiarcherella christellae gen. et sp. nov. The amoebae described herein originate from highly fossiliferous amber pieces. Based on syninclusions, Hemiarcherella christellae was a soil‐dwelling organism, probably an active bacterivore. This taxon represents the third species of testate amoebae described from mid‐Cretaceous French amber. Analysis of this fossil amoeba fauna illustrates the uniqueness of mid‐Cretaceous French amber deposits. Indeed, most amoebae found in amber have been assigned to modern species, corroborating the hypothesis of morphological stasis in different microbial lineages. However, the well‐preserved amoebae fauna found in French amber can be distinguished clearly from modern species and help us to better understand the fossil record of these organisms.     

贵州寒武系第二统杷榔组有壳变形虫的新发现*

有壳变形虫(testate amoebae)的演化历史最早可追溯至新元古代早期, 以该时期北美、华北、挪威以及澳洲等多个地区浅海相碳酸盐岩、页岩中发现的瓶状微体化石(vase-shaped microfossils)为标志。此前认为, 显生宙的有壳变形虫最早出现在早泥盆世。长期以来, 早古生代的地层中未发现这类原生生物的明确化石证据。本研究通过对岩石样品进行常规孢粉酸泡分析处理和切磨岩石薄片, 获取原位保存的化石标本的技术方法, 从贵州东部剑河县交榜剖面出露的寒武系杷榔组(第2统第4阶)中获得数枚有壳变形虫(testate amoebae)化石标本。基于标本的显微形态特征, 并结合激光拉曼光谱等研究, 对原先记述为疑源类的Plagasphaera balangensis和P. sp. A两形态种进行重新认识和描述。由于它们在结构和形态上与一些现生的鳞壳虫目(Euglyphida)有壳变形虫极为相似, 因此将先前定为疑源类的Plagasphaera balangensis和P. sp. A两形态属、种名, 分别修订为Palaeoassulina balangensis gen. et sp. nov.和?Palaeoassulina sp. A。该发现不仅将显生宙有壳变形虫的原有化石记录从晚古生代泥盆纪向前延伸至寒武纪早期, 还为调查研究有壳变形虫的系统演化提供关键的生物化石证据。     

Testate Amoeba (Arcellinida,Euglyphida) Ecology along a Poor‐Rich Gradient in Fens of Western Poland

We investigated the relationships between testate amoebae (Arcellinida, Euglyphida), vegetation and water chemistry along environmental gradients in minerotrophic peatlands (fens) in western Poland. We hypothesized that: a) hydrochemistry significantly influences structure of testate amoeba communities, and b) testate amoeba communities are more closely correlated with the hydrochemical variables (environment) than with the vegetation data. Testate amoeba communities and vegetation from 71 sample plots were investigated together with the hydro‐chemistry and hydrology based on 16 environmental variables and vegetation composition. Testate amoeba communities revealed a distinctive poor‐rich gradient in analysed fens. Mineral‐rich habitats, which were dominated by brown mosses, were preferred by a higher number of taxa than acidic habitats, which were dominated by Sphagnum. We recorded a total of 107 testate amoebae taxa. The average species richness of testate amoebae for brown mosses was higher (20) than for Sphagnum (13). We found that testate amoebae communities were similarly correlated with vascular plants, mosses and environmental parameters. Results of direct ordination demonstrate that hydrology, pH, Mg²⁺ and sodium remain the most important environmental control for the entire data set. CCA showed that in case of brown mosses hydrology, sodium and oxygen affect testate amoeba communities significantly whereas in Sphagnum only sodium emerge as most significant determining testate amoeba assemblages. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

What is the upper size limit for cosmopolitan distribution in free‐living microorganisms?

A comparison of testate amoebae assemblages from the Arctic and Antarctic (areas of similar habitat a maximum distance apart) is used to try and answer the question ‘What is the upper size limit for cosmopolitan distribution in free‐living microbes?’ Species restricted to either the Arctic or Antarctic exhibited sizes up to 230 μm while the largest cosmopolitan species was 135 μm in size. Comparison of the testate assemblages using a multivariate classificatory technique (TWINSPAN ) also suggested more restricted distribution for the larger species. There was a negative relationship between species size and number of sites at which it was recorded (r_s=?0.261, P < 0.05), with all the more widespread species having a size of below 100 μm. It is suggested that for testate amoebae cosmopolitan distributions become common below 100–150 μm. This suggests that most species of testate (indeed most free‐living microbes) have low species richness because of lack of opportunities for allopatric speciation as most are below 100 μm and so geographical isolation is unlikely. It is suggested that if this is correct, only the largest free‐living microbes (> 150 μm) are likely to be of conservation concern because of their smaller ranges. However, I point out that currently different studies are giving very different answers to the question, how ubiquitous and species rich are free‐living microbes? The subject requires further work to try and reconcile these different results.     

Diverse fossil amoebae in German Mesozoic amber

Fossil amoebae are very rare, although their evolutionary history extends back into the Proterozoic. The Cenomanian amber of Schliersee (southern Germany) is very rich in micro-organisms and contains the highest diversity of fossil freshwater rhizopods (Gymnamoebia and Testacealobosia) yet discovered. Altogether seven testate amoebae and one gymnamoebian species are recorded from this Mesozoic amber. The four newly discovered taxa described in this paper can be assigned to the extant species Centropyxis delicatula , Centropyxis hirsuta , Phryganella acropodia and Phryganella paradoxa . Over 200 individuals of Phryganella paradoxa are preserved. Together with their syninclusions, the amoebae are species of limnetic or limnetic–terrestrial microcoenoses. The presence of 100-myr-old fossils with extant representatives suggests evolutionary stasis of these freshwater amoebae. However, not all modern testacean families have been recorded from Mesozoic limnetic habitats. Our experimental studies verify that naked and testate amoebae can be embedded in resins.     

TESTATE AMOEBAE FROM THE EARLY JURASSIC OF THE WESTERN TETHYS,NORTH‐EAST ITALY

Abstract: Fossil testate amoebae and their non‐marine finds are rare so their ecological importance through Earth history is poorly understood. The Lower Jurassic shallow water black‐shales of Trento Platform (north‐east Italy) are rich in micro‐organisms and contain a thecamoebian and ostracod assemblage representing the first known record of Early Jurassic oligohaline forms from the European mainland. The thecamoebians are represented by the genera Difflugia, Pontigulasia and Centropyxis. The present discovery of Lower Jurassic thecamoebians in fine carbonate organic‐rich deposits indicates, for the first time, that these sediments can preserve testate amoebae very well. The occurrence of difflugid testate amoebians confirm a transitional marine‐terrestrial habitat, outside large bodies of water, and suggests occasional eutrophication in ephemeral restricted aquatic environment in the Sinemurian–Pliensbachian Trento Platform.     

Nebela jiuhuensis nov. sp. (Amoebozoa; Arcellinida; Hyalospheniidae): A New Member of the Nebela saccifera ‐ equicalceus ‐ ansata Group Described from Sphagnum Peatlands in South‐Central China

Hyalospheniids are among the most common and conspicuous testate amoebae in high‐latitude peatlands and forest humus. These testate amoebae were widely studied as bioindicators and are increasingly used as models in microbial biogeography. However, data on their diversity and ecology are still very unevenly distributed geographically: notably, data are lacking for low‐latitude peatlands. We describe here a new species, Nebela jiuhuensis, from peatlands near the Middle Yangtze River reach of south‐central China with characteristic morphology. The test (shell) has hollow horn‐like lateral extensions also found in N. saccifera, N. equicalceus (=N. hippocrepis), and N. ansata, three large species restricted mostly to Sphagnum peatlands of Eastern North America. Mitochondrial cytochrome oxidase (COI) data confirm that N. jiuhuensis is closely related to the morphologically very similar North American species N. saccifera and more distantly to N. ansata within the N. penardiana group. These species are all found in wet mosses growing in poor fens. Earlier reports of morphologically similar specimens found in South Korea peatlands suggest that N. jiuhuensis may be distributed in comparable peatlands in Eastern Asia (China and Korea). The discovery of such a conspicuous new species in Chinese peatlands suggests that many new testate amoebae species are yet to be discovered, including potential regional endemics. Furthermore, human activities (e.g., drainage, agriculture, and pollution) have reduced the known habitat of N. jiuhuensis, which can thus be considered as locally endangered. We, therefore, suggest that this very conspicuous micro‐organism with a probably limited geographical distribution and specific habitat requirement should be considered as a flagship species for microbial biogeography as well as local environmental conservation and management.     

The curse of taxonomic uncertainty in biogeographical studies of free-living terrestrial protists: a case study of testate amoebae from Amsterdam Island

Aim A current debate in microbial biogeography contrasts two views concerning the distribution of free‐living microorganisms. The first view assumes a ubiquitous distribution, while the second assumes that at least some species have limited geographical distributions. We tested for limited geographical distributions by identifying testate amoebae morphospecies from an extremely remote oceanic island where the potential for endemism is high. Location Amsterdam Island, Indian Ocean. Methods Sixty moss and water samples collected from the top of the volcano to the lowland were investigated for their testate amoeba content. Due to taxonomic uncertainties among the Argynnia (Nebela) dentistoma species complex (including A. antarctica), we also performed light and scanning electron microscopy investigations on the shell ultrastructure and biometric analyses on several specimens of this taxon. Results We identified a total of 43 testate amoeba taxa belonging to 15 genera. Only four testate amoeba taxa had previously been recorded on this island. Testate amoeba communities of Amsterdam Island are dominated by cosmopolitan ubiquitous euglyphid taxa such as Trinema lineare, Assulina muscorum and Corythion dubium. The length and width ranges for Argynnia dentistoma on Amsterdam Island overlap with other records of this species and of A. antarctica, suggesting that A. antarctica is not a distinct taxon. Main conclusions Although Amsterdam Island is among the most remote islands in the world, an extensive inventory of testate amoeba morphospecies provided no clear evidence for endemism. On the one hand, our detailed morphometric analysis of the A. dentistoma complex revealed that A. antarctica, a morphospecies previously suggested to display endemism, cannot be confidently distinguished from the cosmopolitan morphospecies A. dentistoma. On the other hand, five morphotaxa could not be identified with certainty and might represent new species, potentially with limited distribution. These examples illustrate how taxonomic uncertainties undermine biogeographical studies of testate amoebae. In order to allow better interpretation of morphology‐based testate amoeba distribution data, an assessment of genetic diversity among and within morphotaxa in relation to geographical distance for some common testate amoebae should be given high priority.     

Diverse Legionella‐Like Bacteria Associated with Testate Amoebae of the Genus Arcella (Arcellinida: Amoebozoa)

Diverse species of Legionella and Legionella‐like amoebal pathogens (LLAPs) have been identified as intracellular bacteria in many amoeboid protists. There are, however, other amoeboid groups such as testate amoeba for which we know little about their potential to host such bacteria. In this study, we assessed the occurrence and diversity of Legionella spp. in cultures and environmental isolates of freshwater arcellinid testate amoebae species, Arcella hemispherica, Arcella intermedia, and Arcella vulgaris, via 16S rRNA gene sequence analyses and fluorescent in situ hybridization (FISH). Analysis of the 16S rRNA gene sequences indicated that A. hemispherica, A. intermedia, and A. vulgaris host Legionella‐like bacteria with 94–98% identity to other Legionella spp. based on NCBI BLAST search. Phylogenetic analysis placed Legionella‐like Arcella‐associated bacteria (LLAB) in three different clusters within a tree containing all other members of Legionella and LLAPs. The intracellular localization of the Legionella within Arcella hosts was confirmed using FISH with a Legionella‐specific probe. This study demonstrates that the host range of Legionella and Legionella‐like bacteria in the Amoebozoa extends beyond members of “naked” amoebae species, with members of the testate amoebae potentially serving an ecological role in the dispersal, protection, and replication of Legionella spp. in natural environments.     

Composition of Soil Testate Amoebae Communities: Their Structure and Modifications in the Temperate Rain Forests of New Zealand and Tasmania

A study of the temperate rain forests of New Zealand and Tasmania showed that their soil testate amoebae communities are composed of five groups of taxa: (1) seven taxa characteristic of wet acidic soils and Sphagnum peatlands (i.e., Amphitemidae, Apodera, Alcodera, Certesella, Cyphoderia, Placocista); (2) a group of 16 species of predatory Nebelids and Heleopera spp., characteristic of Sphagnum and rainforests; (3) a group of 17 species of litter and soil Euglypha, excluding the smallest ones; (4) a diverse population of other morphotypes common in other biomes; and (5) a population of small euryoecious taxa – Cryptodifflugia and Pseudodifflugia spp., Euglypha rotunda, E. laevis, Corythion and Trinema spp. This fifth group, with other r‐selected protists (e.g., colpodid ciliates) appears in all habitats. Soil testate communities of other rainforests are composed of the same five groups and are distinguished by the first three assemblages. The fourth and fifth groups, often supplemented with a few Euglypha species, comprise the soil testate amoebae of other biomes. Nebelids and Heleopera, incorporating prey idiosomes into their shells, add an additional link to the role of Euglyphids in the silica cycle. Three Gondwanan Nebelid genera, Apodera, Alcodera, and Certesella were frequently observed, and the discovery of Alcodera cockayni in Tasmania extends its recorded distribution in the Southern Hemisphere.     

Fluctuations of Testate Amoebae Populations (Rhizopoda) in Plankton from Different Environments on the Upper Paraná River Floodplain,Brazil

Fluctuations in temporal and spatial abundance are remarkable features of all animal populations. Regional synchrony of local population dynamics has been verified in several studies. This study investigated the existence of coherent patterns in temporal fluctuations of the principal testate amoebae population abundance in plankton from the upper Paraná river floodplain in Brazil. Testate amoebae sampling was carried out every three months, from February 2000 to November 2002, and twice in 2003. Sampling occurred in 12 environments associated with three different rivers. Samples were obtained at the subsurface of the pelagic region, using a motorized pump and a 68 μm mesh plankton net. Ninety–three testate amoebae species from eight families were identified. The families Difflugiidae, Arcellidae, and Centropyxidae had the greatest number of species. Arcella conica, A. discoides, A. vulgaris, Centropyxis aculeata, C. ecornis, Difflugia gramen, D. pseudogramen and Cucurbitella dentata were the most abundant species, and they were chosen for spatial synchrony analysis. No coherent pattern of abundance fluctuation was found, in any of the floodplain environments studied. Nevertheless, when analyzing only the channels, spatial synchrony was observed to Centropyxis ecornis. Mantel tests showed that the synchrony level, as identified by Spearman and Pearson correlations, was not correlated to geographic distance between environments. We conclude that regional factors in the Upper Paraná River floodplain are not the main determinants of the population dynamics of testate amoebae. The high floodplain heterogeneity seems determine a great relevance of local factors. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Adhesion plaques associated with the production of a daughter cell in Euglypha (Testacea; Potozoa)

Summary Euglypha acanthophora and Euglypha strigosa, testate amoebae with siliceous shells, undergo binary fission producing daughter cells. The siliceous plates from which the shells are constructed are produced in the Golgi and perinuclear regions of the parent. At binary fission these pass along microtubule pathways and are manoeuvered into position in the daughter-cell by microtubule (20–25 nm) and microfilament (7–9 nm) systems. The latter in the form of adhesion plaques are instrumental in the coordination and deployment of the shell-plates.     

Flourish or Flush: Effects of Simulated Extreme Rainfall Events on Sphagnum-dwelling Testate Amoebae in a Subarctic Bog (Abisko, Sweden)

Extreme precipitation events are recognised as important drivers of ecosystem responses to climate change and can considerably affect high-latitude ombrotrophic bogs. Therefore, understanding the relationships between increased rainfall and the biotic components of these ecosystems is necessary for an estimation of climate change impacts. We studied overall effects of increased magnitude, intensity and frequency of rainfall on assemblages of Sphagnum-dwelling testate amoebae in a field climate manipulation experiment located in a relatively dry subarctic bog (Abisko, Sweden). The effects of the treatment were estimated using abundance, species diversity and structure of living and empty shell assemblages of testate amoebae in living and decaying layers of Sphagnum. Our results show that increased rainfall reduced the mean abundance and species richness of living testate amoebae. Besides, the treatment affected species structure of both living and empty shell assemblages, reducing proportions of hydrophilous species. The effects are counterintuitive as increased precipitation-related substrate moisture was expected to have opposite effects on testate amoeba assemblages in relatively dry biotopes. Therefore, we conclude that other rainfall-related factors such as increased infiltration rates and frequency of environmental disturbances can also affect testate amoeba assemblages in Sphagnum and that hydrophilous species are particularly sensitive to variation in these environmental variables.     

The Ecology of Testate Amoebae (Protists) in Sphagnum in North-western Poland in Relation to Peatland Ecology

We studied the relationship between testate amoebae (Protozoa) communities and the depth to the water table (DWT), pH, conductivity, and microhabitat type in Sphagnum dominated peatlands of north-western Poland and built predictive (transfer function) models for inferring DWT and pH based on the testate amoebae community structure. Such models can be used for peatland monitoring and paleoecology. A total of 52 testate amoebae taxa were recorded. In a redundancy analysis, DWT and pH explained 20.1% of the variation in the species data and allowed us to identify three groups of taxa: species that are associated with (1) high DWT and low pH, (2) low DWT and low pH, and (3) high pH and mid-range DWT. Our transfer function models allow DWT and pH to be estimated with mean errors of 9.89 cm and 0.71 pH units. The prediction error of the DWT model and the tolerance of the species both increase with increasing dryness. This pattern mirrors the ecology of Sphagnum mosses: Species growing in wet habitats are more sensitive to change in water table depth than the species growing in drier microhabitats. Our results are consistent with studies of testate amoeba ecology in other regions, and they provide additional support for the use of these organisms in paleoecological and biomonitoring contexts.     

Biogeography of testate amoebae in the soils of Mexico

We studied testate amoebae (testates) in the soils of coniferous and deciduous forests and in the wetland and aquatic habitats of Mexico. In 141 samples we found 205 taxa identified to the species or intraspecies level and 68 testate amoebae, which could not be identified to the species level. The highest species diversity of testates was found in the soils of the tropical rainforest (126 species and intraspecific taxa, including spp.) and in tropical wetlands (144 species and intraspecific taxa, including spp.). The study documented testate amoebae with a limited geographical distribution (genera Centropyxis, Cornuapyxis, Ellipsopyxis, Hoogenraadia, Planhoogenraadia, Apolimia, Certesella, Apodera, and Alocodera). We found that testate amoebae in the soils of Mexico had a high level of polymorphism and individual variability. We discuss the value of soil testate amoebae for defining biogeographical regions of Mexico.     

Grazing on <Emphasis Type="Italic">Microcystis</Emphasis> (Cyanophyceae) by testate amoebae with special reference to cyanobacterial abundance and physiological state

We examined the growth of testate amoebae preying on Microcystis whose physiological states were different in laboratory experiments and a hypertrophic pond. We prepared three experimental systems using water samples dominated by Microcystis aeruginosa: light incubation (control), dark incubation (dark), and light incubation with addition of nitrogen and phosphorus (+NP). In all the systems, the colony density of M. aeruginosa decreased slightly during incubation. Physiological activity of phytoplankton as determined by chlorophyll fluorescence was high and almost constant in the control and +NP systems, whereas it decreased in the dark system. Cell densities of testate amoebae increased in the control and +NP systems, whereas in the dark system they remained low. Thus, growth of the amoebae was low in the systems where physiological activity of Microcystis was low. In a hypertrophic pond, cell density of testate amoebae increased and remained high when M. aeruginosa predominated. Cell density of testate amoebae increased remarkably, simultaneously with the increases in M. aeruginosa colony density and phytoplankton physiological activity. We also found a significant correlation between densities of M. aeruginosa colonies and testate amoebae. We suggested that the physiological activity of Microcystis is one important factor affecting the growth of testate amoebae grazing on Microcystis.     

Seasonal Influences on the Ecology of Testate Amoebae (Protozoa) in a Small <Emphasis Type="Italic">Sphagnum</Emphasis> peatland in Southern Ontario,Canada

Testate amoebae (Protozoa) were studied in spring, summer, and fall from the same microhabitats in a small Sphagnum-dominated peatland in southern Ontario, Canada. A total of 32 sampling stations were established in two wetland plant communities, 19 in an open Ericaceae low-shrub community and 13 in a closed Picea mariana and Larix laricina swamp community. Sphagnum was collected in each station for analysis of testate amoebae and measurement of soil water content parameters and water table depth in May, August, and October 2001. pH and dissolved oxygen of the groundwater under the Sphagnum were measured also. A total of 52 taxa including the rotifer, Habrotrocha angusticollis, were identified. Soil water content and water table variables emerged as the primary factors separating testate amoebae between the open bog/fen community and swamp community. Testate amoebae in the open bog/fen community showed a clear separation between the May sampling period and the August and October sampling periods. Sampling stations in May had much higher water table and were wetter than those in August and October. Conversely, testate amoebae in the swamp community did not show a clear difference between sampling periods. Soil moisture and water tables appear to be more constant in the swamp communities. Biological factors or other microscale environmental factors may need to be considered to explain seasonal changes in testate amoebae. A greater understanding of relationships between testate amoebae and microenvironmental factors is necessary to track seasonality in testate amoebae distributions.     

Sphagnum-dwelling testate amoebae in subarctic bogs are more sensitive to soil warming in the growing season than in winter: the results of eight-year field climate manipulations

Sphagnum-dwelling testate amoebae are widely used in paleoclimate reconstructions as a proxy for climate-induced changes in bogs. However, the sensitivity of proxies to seasonal climate components is an important issue when interpreting proxy records. Here, we studied the effects of summer warming, winter snow addition solely and winter snow addition together with spring warming on testate amoeba assemblages after eight years of experimental field climate manipulations. All manipulations were accomplished using open top chambers in a dry blanket bog located in the sub-Arctic (Abisko, Sweden). We estimated sensitivity of abundance, diversity and assemblage structure of living and empty shell assemblages of testate amoebae in the living and decaying layers of Sphagnum. Our results show that, in a sub-arctic climate, testate amoebae are more sensitive to climate changes in the growing season than in winter. Summer warming reduced species richness and shifted assemblage composition towards predominance of xerophilous species for the living and empty shell assemblages in both layers. The higher soil temperatures during the growing season also decreased abundance of empty shells in both layers hinting at a possible increase in their decomposition rates. Thus, although possible effects of climate changes on preservation of empty shells should always be taken into account, species diversity and structure of testate amoeba assemblages in dry subarctic bogs are sensitive proxies for climatic changes during the growing season.     

The Impact of Simulated Sulfate Deposition on Peatland Testate Amoebae

Peatlands subjected to sulfate deposition have been shown to produce less methane, believed to be due to competitive exclusion of methanogenic archaea by sulfate-reducing bacteria. Here, we address whether sulfate deposition produces impacts on a higher microbial group, the testate amoebae. Sodium sulfate was applied to experimental plots on a Scottish peatland and samples extracted after a period of more than 10 years. Impacts on testate amoebae were tested using redundancy analysis and Mann–Whitney tests. Results showed statistically significant impacts on amoebae communities particularly noted by decreased abundance of Trinema lineare, Corythion dubium, and Euglypha rotunda. As the species most reduced in abundance are all small bacterivores we suggest that our results support the hypothesis of a shift in dominant prokaryotes, although other explanations are possible. Our results demonstrate the sensitivity of peatland microbial communities to sulfate deposition and suggest sulfate may be a potentially important secondary control on testate amoebae communities.     

Testate amoebae and tintinnids as spatial and seasonal indicators in the intertidal margins of Guadiana Estuary (southeastern Portugal)

The present study gathers pioneering taxonomical and community data on testate amoebae and agglutinated tintinnids, analyzing their seasonal and spatial distribution patterns in the mesotidal system of the Guadiana Estuary, southeastern Portugal. To evaluate both groups’ potential as bioindicators in climate monitoring and paleoenvironmental reconstructions, their abundance, diversity and living proportions were compared to elevation in relation to mean sea-level, marine influence and to periods of elevated water levels. The distributions of testate amoebae and tintinnid total assemblages were also related to major physicochemical variables by means of multivariate analysis.From 49 surface sediment samples collected in winter and summer 2010, 17 species (25 strains) of testate amoebae were identified (in the 63 μm fraction). The Centropyxidae, mainly represented by Centropyxis aculeata, Centropyxis arcula and Centropyxis constricta, had the greatest density (71% of total individuals), while the Difflugidae represented the most species (82% of total species). Higher diversities and densities were observed in winter in the upper estuary, where salinity is negligible, and in the middle estuary where they are concentrated at the lower levels of the intertidal zone. Few or no individuals were observed in the highest marsh zones. In winter, dead testate amoebae were dominant, with empty tests accumulating in the sediments from post-mortem transport by high river discharge. In summer, an increase in the living fraction is observed, with living testate amoebae along the entire estuary, which could indicate that some species are able to live in the lower reaches of the estuary.Along with the testate amoebae, two species of agglutinated tintinnids (>63 μm), Stenosemella ventricosa and Tintinnopsis cf. lata, were abundant in the sediments. Their highest abundances were observed in summer. Living individuals were only recorded in summer, mainly in the vicinities of freshwater and sewage outflows, where elevated nutrient concentrations may be expected.The present study demonstrates that both testate amoebae and tintinnids have well defined patterns in their temporal and spatial distribution, offering high bioindicator potential in environmental/climate monitoring studies as well as in paleoenvironmental reconstructions.