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An ever-increasing diversity of potentially toxic chemical compounds are being developed and released into the environment as a result of human activities (e.g. agriculture, drugs, and cosmetics). Among these, pesticides have been shown to affect non-targeted wildlife since the 1960s. A range of ecotoxicological tests are used to assess the toxicity of pesticides on various model organisms. However most model organisms are metazoans, while the majority of Eukaryotes are unicellular microorganisms known as protists. Protists are ubiquitous organisms of key functional roles in all ecosystems but are so far little studied with respect to pesticide impact. To fill this gap, we developed a new ecotoxicological test based on Euglypha rotunda , a common soil amoeba, grown in culture flask with Escherichia coli as sole food source. We tested this assay with the herbicide S -metolachlor, which is known to affect cell division in seedling shoots and roots of weeds. Reproducible growth conditions were obtained for E. rotunda . The growth of E. coli was not affected by the herbicide. The growth of E. rotunda was affected by the herbicide in a non-linear way, growth being significantly reduced at ca. 15 μg/L, but not at 150 μg/L. Our results show the potential for using soil protists in ecotoxicology and adds to the growing body of evidence for non-linear impacts of pesticides on non-target organisms. With the acquisition of additional data, the protocol should be suitable for standard ecotoxicological tests. [ABSTRACT FROM AUTHOR]

Protists are the most diverse eukaryotes. These microbes are keystone organisms of soil ecosystems and regulate essential processes of soil fertility such as nutrient cycling and plant growth. Despite this, protists have received little scientific attention, especially compared to bacteria, fungi and nematodes in soil studies. Recent methodological advances, particularly in molecular biology techniques, have made the study of soil protists more accessible, and have created a resurgence of interest in soil protistology. This ongoing revolution now enables comprehensive investigations of the structure and functioning of soil protist communities, paving the way to a new era in soil biology. Instead of providing an exhaustive review, we provide a synthesis of research gaps that should be prioritized in future studies of soil protistology to guide this rapidly developing research area. Based on a synthesis of expert opinion we propose 30 key questions covering a broad range of topics including evolution, phylogenetics, functional ecology, macroecology, paleoecology, and methodologies. These questions highlight a diversity of topics that will establish soil protistology as a hub discipline connecting different fundamental and applied fields such as ecology, biogeography, evolution, plant-microbe interactions, agronomy, and conservation biology. We are convinced that soil protistology has the potential to be one of the most exciting frontiers in biology. [ABSTRACT FROM AUTHOR]

Soil bacterial community structures and associated ecosystem functions are known to vary both in space and time but spatial and temporal patterns are rarely studied simultaneously. Dynamic floodplains offer an ideal setting to study these spatio-temporal patterns and to determine the relative importance of habitat diversity and seasons in shaping soil bacterial communities. We studied the differences between habitats and seasonal patterns of soil bacterial communities (T-RFLPs) and soil environmental conditions (temperature, moisture and time elapsed since last inundation) and ecosystem functioning proxies (basal respiration, enzymatic activity, microbial C and N) over one year in seven habitats of a restored floodplain. Soil bacterial community structures in the two habitats closest to the river were most divergent from the others. However, although bacterial community structures differed between habitats at all sampling times, their seasonal variability was substantially higher than the habitat differences. Most of the ecosystem functioning proxies were lowest in the most dynamic habitat (gravel bar). Furthermore, while both bacterial community structure and ecosystem functions varied in relation to environmental conditions, the two were not directly correlated, and the link between communities and functions was seasonally inconsistent. The ratio of seasonal vs. habitat variability of soil bacterial communities decreased from the most dynamic to the most stable (forest) habitat. Thus in dynamic ecosystems such as floodplains also external factors (flood disturbance, seasonally changing climatic conditions) might influence ecosystem functions and bacterial communities independently – possibly because of species functional redundancy – and are keys in maintaining floodplain taxonomical and functional diversity. [ABSTRACT FROM AUTHOR]

Abstract: Sound taxonomy is a major requirement for quantitative environmental reconstruction using biological data. Transfer function performance should theoretically be expected to decrease with reduced taxonomic resolution. However for many groups of organisms taxonomy is imperfect and species level identification not always possible. We conducted numerical experiments on five testate amoeba water table (DWT) transfer function data sets. We sequentially reduced the number of taxonomic groups by successively merging morphologically similar species and removing inconspicuous species. We then assessed how these changes affected model performance and palaeoenvironmental reconstruction using two fossil data sets. Model performance decreased with decreasing taxonomic resolution, but this had only limited effects on patterns of inferred DWT, at least to detect major dry/wet shifts. Higher-resolution taxonomy may however still be useful to detect more subtle changes, or for reconstructed shifts to be significant. [Copyright &y& Elsevier]

Abstract: Peatlands are widely exploited archives of paleoenvironmental change. We developed and compared multiple transfer functions to infer peatland depth to the water table (DWT) and pH based on testate amoeba (percentages, or presence/absence), bryophyte presence/absence, and vascular plant presence/absence data from sub-alpine peatlands in the SE Swiss Alps in order to 1) compare the performance of single-proxy vs. multi-proxy models and 2) assess the performance of presence/absence models. Bootstrapping cross-validation showing the best performing single-proxy transfer functions for both DWT and pH were those based on bryophytes. The best performing transfer functions overall for DWT were those based on combined testate amoebae percentages, bryophytes and vascular plants; and, for pH, those based on testate amoebae and bryophytes. The comparison of DWT and pH inferred from testate amoeba percentages and presence/absence data showed similar general patterns but differences in the magnitude and timing of some shifts. These results show new directions for paleoenvironmental research, 1) suggesting that it is possible to build good-performing transfer functions using presence/absence data, although with some loss of accuracy, and 2) supporting the idea that multi-proxy inference models may improve paleoecological reconstruction. The performance of multi-proxy and single-proxy transfer functions should be further compared in paleoecological data. [Copyright &y& Elsevier]

Abstract: Peatland testate amoebae are widely used to reconstruct paleohydrological/climatic changes, but many species are also known to respond to pollutants. Peatlands around the world have been exposed to anthropogenic and intermittent natural pollution through the late Holocene. This raises the question: can pollution lead to changes in the testate amoeba paleoecological record that could be erroneously interpreted as a climatic change? To address this issue we applied testate amoeba transfer functions to the results of experiments adding pollutants (N, P, S, Pb, O3) to peatlands and similar ecosystems. We found a significant effect in only one case, an experiment in which N and P were added, suggesting that pollution-induced biases are limited. However, we caution researchers to be aware of this possibility when interpreting paleoecological records. Studies characterising the paleoecological response to pollution allow pollution impacts to be tracked and distinguished from climate change. [Copyright &y& Elsevier]

A massive tufa complex in the lower Lorze Valley (550 m a.s.l., north-central Switzerland) formed between 9000 and 5000 cal. yr BP. In the plain 4 km southeast of the complex lies Neugrundmoor (830 m a.s.l.), a mire in the hydrologic recharge area of the springs responsible for the tufa formation. Here we address an earlier, untested hypothesis that the transition of Neugrundmoor from fen to raised bog caused the end of the tufa formation, by means of pollen and testate amoebae analyses on a peat section. The chronology of the section is based on ten AMS 14C-dates of terrestrial plant macrofossils. Our results suggest that until 7100 cal. yr BP the plain had shallow ponds, which gradually developed into fens. Starting 5350 cal. yr BP, large raised bogs developed in the fens in a few centuries. This agrees with the hypothesis that the end of tufa formation coincided with the transition from fen to raised bog. It also coincided with the end of the mid-Holocene temperature optimum, when climate became cooler and wetter. We suggest that the main factor causing the end of tufa formation at 5500-5000 cal. yr BP was declining air temperatures, which on the one hand led to diminished calcium precipitation, on the other to enhanced bog formation. Peat has a low hydraulic conductivity, so we suggest that less water became available for tufa formation because of increased thickness of peat layers, exacerbating the direct effect of cooling on calcium precipitation. Noticeable human influence in the surroundings did presumably not start before c. 3500 cal. yr BP and was then very weak, so this was not responsible for the end of the tufa formation. Extrapolation of our results suggests that climatic cooling alone is sufficient to explain the widespread termination of tufa formation after the mid Holocene. [ABSTRACT FROM AUTHOR]

Abstract: Marine and freshwater ecosystems are fundamentally different regarding many biotic and abiotic factors. The physiological adaptations required for an organism to pass the salinity barrier are considerable. Many eukaryotic lineages are restricted to either freshwater or marine environments. Molecular phylogenetic analyses generally demonstrate that freshwater species and marine species segregate into different sub-clades, indicating that transitions between these two environments occur only rarely in the course of evolution. It is, however, unclear if the transitions between freshwater and environments characterized by highly variable salinities, such as the marine supralittoral zone, are also infrequent. Here, we use testate amoebae within the Euglyphida to assess the phylogenetic interrelationships between marine supralittoral and freshwater taxa. Euglyphid testate amoebae are mainly present in freshwater habitats but also occur in marine supralittoral environments. Accordingly, we generated and analyzed partial SSU rRNA gene sequences from 49 new marine/supralittoral and freshwater Cyphoderiidae sequences, 20 sequences of the Paulinellidae, Trinematidae, Assulinidae, and Euglyphidae families as well as 21 GenBank sequences of unidentified taxa derived from environmental PCR surveys. Both the molecular and morphological data suggest that the diversity of Cyphoderiidae is strongly underestimated. The results of our phylogenetic analyses demonstrated that marine supralittoral and freshwater euglyphid testate amoeba species are segregated into distinct sub-clades, suggesting that transitions between these two habitats occurred only infrequently. [Copyright &y& Elsevier]

Synthetic pesticides such as neonicotinoids are commonly used to treat crops in tropical regions, where data on environmental and human contamination are patchy and make it difficult to assess to what extent pesticides may harm human health, especially in less developed countries. To assess the degree of environmental and human contamination with neonicotinoids we collected soil, water and people's hair in three agricultural regions of the Philippines and analysed them by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS-MS). Five neonicotinoids, namely acetamiprid, clothianidin, imidacloprid, thiacloprid and thiamethoxam were targeted. Residues of neonicotinoids were found in 78% of 67 soil samples from the three provinces. Total neonicotinoid loads ranged on average between 0.017 and 0.89 μg/kg in soils of rice, banana and vegetable crops, and were 130 times higher (113.5 μg/kg) in soils of a citrus grove. Imidacloprid was the most prevalent compound at an average of 0.56 μg/kg in soil while thiacloprid was below the limit of detection. Half of the eight water samples from a rice field and nearby creek contained residues of imidacloprid (mean 1.29 ng/L) and one contained thiamethoxam (0.15 ng/L). Residues of neonicotinoids were found in 81% of 99 samples of people's hair from the surveyed regions (average total concentrations 0.14 to 1.18 ng/g, maximum 350 ng/g). Hair residue levels correlated well with the concentrations of thiamethoxam and total residues in soils from the same locality (r = 0.98). The presence of thiacloprid in 15% of the hair samples but not in soil samples suggests an additional route of exposure among people, which is most likely to be through ingestion of agricultural food and drinks available in the market. Unlabelled Image • Neonicotinoid residues found in soil, water and people's hair in the Philippines • Average residue loads in soil were highest in citrus groves of Mindanao. • Imidacloprid was the most prevalent neonicotinoid found in soil and water. • Thiamethoxam and imidacloprid were most prevalent in hair samples. • Residues in hair correlated well with those in soil from the same localities [ABSTRACT FROM AUTHOR]

Biomonitoring tools are useful to assess the impact of environmental changes on the functioning of ecosystems. Existing tools mostly require species identification, thus allowing to estimating changes in biodiversity, and possibly inferring ecosystem functioning, using functional diversity and traits based approaches. Testate amoebae are good indicators of surface moisture conditions in Sphagnum peatlands and are routinely used in palaeoecology. Their shells (tests), on which identification is based, can also be used to define functional traits and thus to infer changes in ecosystem functioning. We investigated the response of testate amoeba communities to manipulated water table depth (wet: −4 cm, intermediate: −15 cm, and dry: −25 cm) over time (seven time points, 19 months) using mesocosms by comparing two approaches: community structure and functional traits responses, using a combination of morphological (biovolume, length, aperture size and position) and physiological (mixotrophy/heterotrophy, shell material) traits. This is the first study investigating the effect of water table depth on testate amoeba assemblages over time using a mesocosm approach. Taxonomical and functional approaches showed similar response patterns, confirming that water level acted as a strong environmental filter. After one year Hyalosphenia papilio decreased in the dry treatment, and the community structure shifted towards a dominance of dry indicators ( Nebela tincta complex, Corythion dubium, Euglypha compressa ) and the selected functional traits (smaller, heterotrophic, compressed species, with a ventral aperture) corresponded to drought adaptations. In line with recent observational and transfer function studies exploring the use of testate amoebae functional traits, our experimental results illustrate how well-selected traits could be used to monitor the impact of present and past climatic changes on Sphagnum peatlands. [ABSTRACT FROM AUTHOR]

Monitoring tools are needed to assess changes in peatland biotic communities and ecosystem functions in response to on-going climate and other environmental changes. Although the responses of soil organisms and plants to ecological gradients and perturbations do not always correlate, peatland monitoring is mainly based on vegetation surveys. Testate amoebae, a group of protists, are important contributors to carbon and nitrogen cycling in organic soils and are useful bioindicators in peatland ecology and paleoecology. There is however little comparative data on the value of testate amoebae, vascular plants and bryophytes as bioindicators of micro-environmental gradients in peatlands. We compared the relationships of testate amoebae, bryophytes, and vascular plants with soil temperature, water table depth, micro-habitats and the carbon and nitrogen content of Sphagnum mosses in four peatlands along a 1300 m altitudinal gradient in Switzerland. We used the full diversity of vascular plants and bryophyte but only a selection of ten easily identifiable testate amoeba morpho-taxa (i.e. species or species-complexes). Indirect and direct gradient ordinations, multiple factor analysis (MFA) and transfer function models for inferring water table depth showed that a selection of ten testate amoeba taxa are more powerful (% variance explained in RDA) and accurate (discrimination among habitats) indicators of local conditions (micro-habitat type, water table depth and Sphagnum C/N ratio) than the vegetation (vascular plants and bryophytes either individually or combined and considering the full diversity). Our study showed that a limited list of ten easily identifiable testate amoeba taxa have higher bioindication value than the full bryophytes and vascular plants. Furthermore, testate amoebae can be analyzed on samples collected at any season (accessibility allowing and if precise sampling sites are well marked) – a clear advantage for biomonitoring and can be used to infer past changes from the peat record at the same taxonomic resolution. This simple approach could therefore be very useful for biomonitoring of peatlands. [ABSTRACT FROM AUTHOR]

Abstract: Testate amoebae (Arcellinida and Euglyphida) and diatoms (Bacillariophyta) respond to different ecological gradients. These protists are useful tools for biomonitoring and paleoecological studies in aquatic and terrestrial ecosystems. However, little is known about the responses of these microeukaryotes to soil management practices. We analyzed the testate amoeba and diatom communities from the DOK-trial (D: biodynamic, O: bio-organic, K: german ‘‘konventionell’’ integrated conventional) agricultural experiment at Therwil, Switzerland. Soil samples were collected from biodynamic and conventional plots and subsequently incubated for four months in a growth chamber. The diatom diversity tended to be higher in the biodynamic than in the two conventional systems. Redundancy analysis (RDA) suggested that diatom community structure differed between organic and the two conventional systems. Testate amoeba abundance was about five times higher in biodynamic than in conventional systems (P < 0.05) but no significant differences in diversity were reported between treatments. Altogether, these data suggest that diatoms and testate amoebae are sensitive to farming systems. As direct analyses of soil samples are time-consuming, molecular tools would be very useful for further development of the use of protists in bioindication. [Copyright &y& Elsevier]

The peat archives are one of the stratigraphic records revealing clearly physical, chemical and biological signals of human influence on the Earth System since the 1950s, at least. The presented study was aimed mainly to identify the level and origin of anthropogenic radionuclides such as 238, 239, 240Pu in a 210Pb-dated peat profile derived from the Northern Ural, Russian Federation. As stated, the vertical variability of 240Pu/239Pu isotopic compositions reflects the nuclear weapons testing history with the maximum in the 1960s and small regional impact most likely of high-yielded tests in the 1950s as well as Chinese detonations in the 1970s. Peat accumulations rates were similar to those obtained in adjacent areas, whereas 210Pb flux slightly exceeded the reference level established for adequate latitude belt. • Study on Pu isotopes accumulated in peat archive of the Northern Ural, Russia. • Application of CF/CS and CF models based on 210Pb chronology. • As high as 0.43 ± 0.02 of 240Pu/239Pu ratio recorded in the 1950s. • Evidence of the regional impact of Chinese nuclear tests in the 1970s. [ABSTRACT FROM AUTHOR]

Abstract: We reconstructed the Holocene developmental history of a kettle-hole peatland in the Tuchola Forest of Northern Poland, using pollen, testat amoebae and plant macrofossil indicators. Our aims were to determine the timing and pattern of autogenic succession and natural and anthropogenic influences on the peatland. Northern Poland is under mixed oceanic and continental climatic influences but has so far been less studied in a palaeoecological context than more oceanic regions of Europe. In the first terrestrial developmental phase of the mire, the testate amoebae-inferred depth to water table revealed two major dry shifts at ca. 9400 (end of lake phase) and ca. 7100 cal BP (a period of global cooling and dry shift in Western Europe). Conditions became wetter again in two steps at ca. 6700 and ca. 5800 BP after a dry event at ca. 6100 BP. The timing of the wet shift at 5800 BP corresponds to wet periods in Western Europe. Peat accumulation rates were low (0.1 mm yr−1) between ca. 5600 and ca. 3000 BP when sedges dominated the peatland. In the last 2500 yrs surface moisture fluctuated with wet events at ca. 2750–2400, and 2000 BP, and dry events at ca. 2250–2100 and 1450 BP. After 1450 BP a trend towards wetter conditions culminated at ca. 500 cal BP, possibly caused by local deforestation. Over the mire history, pH (inferred from testate amoebae) was mostly low (around 5) with two short-lived shifts to alkaline conditions (7.5) at ca. 6100 and 1450 BP indicating a minerotrophic influence from surface run-off into the mire. Up to about 1000 BP the ecological shifts inferred from the three proxies agree with palaeoclimatic records from Poland and Western Europe. After this date, however correlation is less clear suggesting an increasing local anthropogenic impact on the mire. This study confirms that kettle-hole peatlands can yield useful palaeoenvironmental data as well as recording land-use change and calls for more comparable studies in regions are the interface between major climate influences. [Copyright &y& Elsevier]

In the last two centuries, a high proportion of peatlands have been lost or severely degraded across the world. The value of peatlands is now well-recognised for biodiversity conservation, flood management, and carbon mitigation, with peatland restoration now central to many government policies for climate action. A challenge, however, is to determine 'natural' and 'disturbed' conditions of peatlands to establish realistic baselines for assessing degradation and setting restoration targets. This requires a tool or set of tools that can rapidly and reliably capture peatland condition across space and time. Our aim was to develop such a tool based on combined analysis of plant and testate amoebae; a group of shelled protists commonly used as indicators of ecological change in peatlands. The value of testate amoebae is well established in Northern Hemisphere Sphagnum -dominated peatlands; however, relatively little work has been undertaken for Southern Hemisphere peat forming systems. Here we provide the first assessment and comparison of the bioindicator value of testate amoebae and vascular plants in the context of Southern Hemisphere peatlands. Our results further demonstrate the unique ecohydrological dynamics at play in New Zealand peat forming systems that set them apart from Northern Hemisphere peatlands. Our results show that plant and testate amoeba communities provided valuable information on peatland condition at different scales, we found that testate amoebae tracked changes in the abiotic variables (depth to water table, pH, and conductivity) more closely than vascular plants. Our results further demonstrate that functional traits of testate amoebae showed promising relationships with disturbance. Amoeba test compression, aperture position and test size were linked to changes in hydrology driven by fluctuations in ground water tables; however, trait responses manifested differently in ombrotrophic and minerotrophic peatlands. Overall, testate amoebae provide a promising bioindicator for tracking degradation in New Zealand peatlands and a potential additional tool to assess peatland condition. • The first assessment and comparison of the bioindicator value of testate amoebae and vascular plants in Southern Hemisphere peatlands. • Testate amoebae are a promising bioindicator for tracking degradation in New Zealand peatlands and are a potential tool for peatland managers. • Testate amoebae tracked changes in the abiotic variables more closely than vascular plants in New Zealand peatlands. • Functional trait analysis of testate amoebae enriches our understanding of microorganism response to degradation in peatlands. [ABSTRACT FROM AUTHOR]

Abstract Fossil testate amoeba assemblages have been used to reconstruct peatland palaeohydrology for more than two decades. While transfer function training sets are typically of local-to regional-scale in extent, combining those data to cover broad ecohydrological gradients, from the regional-to continental- and hemispheric-scales, is useful to assess if ecological optima of species vary geographically and therefore may have also varied over time. Continental-scale transfer functions can also maximise modern analogue quality without losing reconstructive skill, providing the opportunity to contextualise understanding of purely statistical outputs with greater insight into the biogeography of organisms. Here, we compiled, at moderate taxonomic resolution, a dataset of nearly 2000 modern surface peatland testate amoeba samples from 137 peatlands throughout North America. We developed transfer functions using four model types, tested them statistically and applied them to independent palaeoenvironmental data. By subdividing the dataset into eco-regions, we examined biogeographical patterns of hydrological optima and species distribution across North America. We combined our new dataset with data from Europe to create a combined transfer function. The performance of our North-American transfer function was equivalent to published models and reconstructions were comparable to those developed using regional training sets. The new model can therefore be used as an effective tool to reconstruct peatland palaeohydrology throughout the North American continent. Some eco-regions exhibited lower taxonomic diversity and some key indicator taxa had restricted ranges. However, these patterns occurred against a background of general cosmopolitanism, at the moderate taxonomic resolution used. Likely biogeographical patterns at higher taxonomic resolution therefore do not affect transfer function performance. Output from the combined North American and European model suggested that any geographical limit of scale beyond which further compilation of peatland testate amoeba data would not be valid has not yet been reached, therefore advocating the potential for a Holarctic synthesis of peatland testate amoeba data. Extending data synthesis to the tropics and the Southern Hemisphere would be more challenging due to higher regional endemism in those areas. Highlights • Dataset of ca. 2000 North American peatland testate amoeba surface samples compiled. • Palaeohydrological transfer functions tested statistically, applied to independent palaeo data. • New model an effective tool for palaeohydrological reconstruction across North America. • Biogeographical patterns occur at taxonomic resolution applied; taxa hydrological optima mostly robust across North America. • Combined North American and European dataset illustrates potential for Holarctic synthesis. [ABSTRACT FROM AUTHOR]

Soil microbial communities are vital for multiple ecosystem processes and services. In particular, soil microbial food webs are key determinants of soil biodiversity, functioning and stability. Unclear, however, is how structural features of food webs, such as species richness and turnover, biomass and energy transfer across trophic levels, influence the provisioning and stability of ecosystem functioning. Here, we explore the relationships between different facets of microbial food web structure (e.g. species richness, connectance, biomass and energy fluxes across trophic levels) and ecosystem functions (i.e. decomposition and microbial enzyme activity) across different habitats and depths in a peatland. We show that no aspect of taxonomic richness directly explained variation in ecosystem functions. Instead, we find that trophic interactions between basal species and primary consumers, and especially increasing connectance, biomass and energy flux transiting from decomposers and phototrophs to algivores, bacterivores and fungivores, enhance ecosystem functions in the peatland. These findings demonstrate that focusing on taxonomic diversity without explicit inclusion of food web structure and energy flows therein gives an incomplete and uninformative comprehension of relationships between biodiversity and ecosystem functioning, at least in peatlands. Our findings further suggest that the inclusion of soil microbial food webs in large-scale biogeochemical models is of fundamental importance to provide the necessary guidance for managing and mitigating the effects of environmental change. • Soil microbial food webs determine soil biodiversity and ecosystem functioning. • What aspect of microbial food web underpin ecosystem functions is unclear. • Biomass and energy flux dynamics, but not taxonomic richness, influence peatland ecosystem functions. • The inclusion of microbial food web properties in biogeochemical models is fundamental. [ABSTRACT FROM AUTHOR]

Sphagnum riparium (Ångström) is a rare constituent of modern peatland plant communities and is also very rarely found as a subfossil in peat archives. We present new data on the occurrence of Sphagnum riparium macrofossils in three Northern Hemisphere peatlands from Yellowknife (NW Canada), Abisko (N Sweden), and the Northern Ural Mountains (NW Russia). Sphagnum riparium macrofossils were present in transitional phases between rich fen and oligotrophic bog. Sphagnum riparium was a dominant species in the three sites and was found in combination with Sphagnum angustifolium , Drepanocladus sp., and vascular plants including Andromeda polifolia, Chamedaphne calyculata and Oxycoccus palustris . Testate amoebae indicate that the species occurred in wet to moderately wet conditions (water-table depth inferred from a testate amoeba transfer function model ranged between 25 and 0 cm under the peatland surface). The wet-indicator taxa Archerella flavum and Hyalosphenia papilio dominated the testate amoeba communities in peat horizons containing Sphagnum riparium. The presence of Sphagnum riparium macrofossils in peat profiles in the Northern Hemisphere can be interpreted as an indication of wet minerotrophic conditions, often corresponding to a rise in water-level and establishment of a wet habitat. Sphagnum riparium is a transient species in these peatlands and is replaced by communities dominated by more acidophilic species such as Sphagnum angustifolium, Sphagnum russowii , and Sphagnum fuscum . Our data show that although Sphagnum riparium is a transient peat-forming species, it is widespread in sub-arctic and boreal environments. The subfossil occurrence of Sphagnum riparium in the Northern Hemisphere may indicate that its range has increased during the Late Holocene. The conservation of Sphagnum riparium in peatlands depends on the existence of relatively short-lived transitional communities which potentially can be artificially created. [ABSTRACT FROM AUTHOR]

Decomposing vertebrate cadavers release large, localized inputs of nutrients. These temporally limited resource patches affect nutrient cycling and soil organisms. The impact of decomposing cadavers on soil chemistry is relevant to soil biology, as a natural disturbance, and forensic science, to estimate the postmortem interval. However, cadaver impacts on soils are rarely studied, making it difficult to identify common patterns. We investigated the effects of decomposing pig cadavers (Sus scrofa domesticus) on soil chemistry (pH, ammonium, nitrate, nitrogen, phosphorous, potassium and carbon) over a one-year period in a spruce-dominant forest. Four treatments were applied, each with five replicates: two treatments including pig cadavers (placed on the ground and hung one metre above ground) and two controls (bare soil and bags filled with soil placed on the ground i.e. "fake pig" treatment). In the first two months (15-59 days after the start of the experiment), cadavers caused significant increases of ammonium, nitrogen, phosphorous and potassium (p<0.05) whereas nitrate significantly increased towards the end of the study (263-367 days; p<0.05). Soil pH increased significantly at first and then decreased significantly at the end of the experiment. After one year, some markers returned to basal levels (i.e. not significantly different from control plots), whereas others were still significantly different. Based on these response patterns and in comparison with previous studies, we define three categories of chemical markers that may have the potential to date the time since death: early peak markers (EPM), late peak markers (LPM) and late decrease markers (LDM). The marker categories will enhance our understanding of soil processes and can be highly useful when changes in soil chemistry are related to changes in the composition of soil organism communities. For actual casework further studies and more data are necessary to refine the marker categories along a more precise timeline and to develop a method that can be used in court. [ABSTRACT FROM AUTHOR]

High-throughput sequencing (HTS) of soil environmental DNA (eDNA) allows assessing the full diversity of soil micro-eukaryotes. The resulting operational taxonomic units (OTUs) can be assigned to potential taxonomic and functional identities using increasingly complete reference databases. HTS of soil eDNA is revealing a high diversity and abundance of potential eukaryovorous protists, thus challenging the paradigm of the predominantly bacterivorous function of soil phagotrophic protists (i.e. microbial loop). Using Illumina sequencing of soil eDNA and targeting the V9 region of the SSU rRNA gene, we investigated the taxonomic and functional diversities, distribution and co-occurrence patterns of soil micro-eukaryotes in three land-use categories: forests, meadows and croplands located in Switzerland. Each OTU was assigned to a broad functional category (phototrophs, phagotrophs, osmotrophs, or parasites). Total OTU richness was similar in the three land-use categories, but community composition differed significantly between forests and other land-uses. The proportion of fungal sequences (especially Basidiomycota) was highest, and phototroph (i.e. soil microalgae) sequences least abundant in forests. Seven OTUs representing phagotrophic protists, together accounting for >25% of all phagotroph sequences, were significantly correlated to the total number of phototroph sequences, thus suggesting algivory. At least three of these OTUs corresponded to known algal predators. These results suggest that beyond plants, soil microalgae represent a functionally significant but rarely considered input of carbon in soils that should be taken into account when modelling soil nutrient cycling. [ABSTRACT FROM AUTHOR]

In terrestrial ecosystems decomposing cadavers act as resource patches affecting nutrient cycling and soil communities, but the effects on soil communities are not well known. In this study we investigated nematode community response to decomposing pig cadavers ( Sus scrofa ) over a one-year period. As nematodes play key roles in soil food webs and are known to respond to disturbances and nutrient enrichment, we hypothesised that they would respond to decomposing cadavers and that this response would change over time. We compared the temporal patterns of nematode density and community structure under pig cadavers, either placed directly on the ground or hung 1 m aboveground (for effects of cadaveric fluids only), with two controls, i.e., bare soil and bags filled with soil placed on the ground (fake pigs – for microclimatic effects only). In the control and fake pig treatments nematode densities, community patterns and maturity indices did not change significantly. In contrast, density increased significantly underneath the ground and hanging pigs two weeks after the beginning of the experiment, and nematode family richness, Simpson diversity and maturity index were significantly reduced in the cadaver treatments. Most nematode families responded negatively to cadavers with the notable exceptions of Rhabditidae, Neodiplogasteridae and Diplogasteroididae. The latter two were found exclusively underneath the decomposing cadavers and are promising bioindicators of vertebrate cadaver decomposition. Even though diversity, density and communities were recovering after one year, the impact of cadavers was still significant for the maturity index. These contrasting patterns illustrate how decomposing cadavers contribute to increasing local biodiversity and suggest that soil nematodes could be used as a tool to document the presence of a decomposing cadaver, or to estimate the time elapsed since death (post-mortem interval). Patterns should, however, be compared in different settings and seasons before such a tool can be validated. [ABSTRACT FROM AUTHOR]

In the decade since the first pan-European testate amoeba-based transfer function for peatland palaeohydrological reconstruction was published, a vast amount of additional data collection has been undertaken by the research community. Here, we expand the pan-European dataset from 128 to 1799 samples, spanning 35° of latitude and 55° of longitude. After the development of a new taxonomic scheme to permit compilation of data from a wide range of contributors and the removal of samples with high pH values, we developed ecological transfer functions using a range of model types and a dataset of ∼1300 samples. We rigorously tested the efficacy of these models using both statistical validation and independent test sets with associated instrumental data. Model performance measured by statistical indicators was comparable to other published models. Comparison to test sets showed that taxonomic resolution did not impair model performance and that the new pan-European model can therefore be used as an effective tool for palaeohydrological reconstruction. Our results question the efficacy of relying on statistical validation of transfer functions alone and support a multi-faceted approach to the assessment of new models. We substantiated recent advice that model outputs should be standardised and presented as residual values in order to focus interpretation on secure directional shifts, avoiding potentially inaccurate conclusions relating to specific water-table depths. The extent and diversity of the dataset highlighted that, at the taxonomic resolution applied, a majority of taxa had broad geographic distributions, though some morphotypes appeared to have restricted ranges. [ABSTRACT FROM AUTHOR]

Transfer functions are valuable tools in palaeoecology, but their output may not always be meaningful. A recently-developed statistical test (‘randomTF’) offers the potential to distinguish among reconstructions which are more likely to be useful, and those less so. We applied this test to a large number of reconstructions of peatland water table depth based on testate amoebae. Contrary to our expectations, a substantial majority (25 of 30) of these reconstructions gave non-significant results (P > 0.05). The underlying reasons for this outcome are unclear. We found no significant correlation between randomTF P-value and transfer function performance, the properties of the training set and reconstruction, or measures of transfer function fit. These results give cause for concern but we believe it would be extremely premature to discount the results of non-significant reconstructions. We stress the need for more critical assessment of transfer function output, replication of results and ecologically-informed interpretation of palaeoecological data. [ABSTRACT FROM AUTHOR]

Annelids (Lumbricidae and Enchytraeidae) and nematodes are common soil organisms and play important roles in organic matter decomposition, nutrient cycling and creation of soil structure and porosity. However, these three groups have rarely been studied together and only few studies exist for urban soils. We studied the diversity and community composition of annelids and nematodes in soils spanning more than two centuries of urban soil development in Neuchâtel (Switzerland) and assessed the relationships 1) among these three groups and 2) between each group and environmental (physical, chemical and functional) characteristics of soils and soil age. While the groups of environmental variables were correlated (Mantel tests) no correlation was found between pairs of soil fauna groups and between each soil fauna group and environmental variables. More specifically, redundancy analyses showed that earthworm assemblages were best correlated with soil bulk density and with soil depth, the latter being positively correlated with soil age. Enchytraeid assemblages and the proportion of enchytraeid r -strategists were respectively best correlated with soil carbonate content and negatively correlated with soil age. Nematodes assemblages were best correlated with soil water content. Moreover, relationships between pairs of soil biota groups, and between each group and environmental (physical, chemical and functional) variables, varied along the soil age gradient (moving window analysis). This study provides new knowledge on urban soil biodiversity and how environmental conditions can influence soil diversity and community patterns in the urban context. The contrasted community patterns of earthworms, enchytraeids and nematodes in urban soils of different ages and their different ecological roles suggest that they represent potential complementary indicators of soil quality and functioning such as soil formation and organic matter dynamics. [ABSTRACT FROM AUTHOR]

Question A better understanding of the response of Sphagnum mosses and associated vascular plants to climate warming is relevant for predicting the carbon balance of peatlands in a warmer world. Open-top chambers ( OTCs) have been used to investigate the effect on soil biogeochemical processes in peatlands, but little information is available on the effects of OTCs on microclimate conditions and the associated response of the plant community. We aimed to understand how simulated warming and differences in soil moisture affect plant species cover. Location A Sphagnum-dominated peatlands in French Jura. Methods We used OTCs to measure the effect of a near-ground temperature increase (+1.5 °C on average) on vegetation dynamics over five growing seasons (2008-2012) in a Sphagnum-dominated peatland, in two adjacent microhabitats with different hydrological conditions - wet and dry. Microclimatic conditions and plant species abundance were monitored at peak biomass in years 1, 2, 3 and 5 and monthly during the plant growing season in year 5. Results The response to warming differed between vascular plants and bryophytes, as well as among species within these groups, and also varied in relation to soil moisture. Andromeda polifolia abundance responded positively to warming, while Vaccinium oxycoccus responded negatively, and Eriophorum vaginatum showed a high resistance. Conclusion Depth of rooting of vascular plants appeared to control the response in plant abundance, while moss abundance depended on various other interacting factors, such as shading by the vascular plant community, precipitation and soil moisture. [ABSTRACT FROM AUTHOR]

Although widely used in ecology, comparative analyses of diversity and niche properties are still lacking for microorganisms, especially focusing on niche variations. Quantifying the niches of microbial taxa is necessary to then forecast how taxa and the communities they compose might respond to environmental changes. In this study, we first identified important topoclimatic, edaphic, spatial and biotic drivers of the alpha and beta diversity of bacterial, archaeal, fungal and protist communities. Then, we calculated the niche breadth and position of each taxon along the important environmental gradients to determine how these vary within and among the taxonomic groups. We found that edaphic properties were the most important drivers of both, community diversity and composition, for all microbial groups. Protists and bacteria presented the largest niche breadths on average, followed by archaea, with fungi displaying the smallest. Niche breadth generally decreased towards environmental extremes, especially along edaphic gradients, suggesting increased specialization of microbial taxa in highly selective environments. Overall, we showed that microorganisms have well defined niches, as do macro-organisms, likely driving part of the observed spatial patterns of community variations. Assessing niche variation more widely in microbial ecology should open new perspectives, especially to tackle global change effects on microbes. • We studied bacterial, archaeal, fungal and protist communities in Alpine soils. • Edaphic properties, particularly pH, drive alpha and beta diversity of all groups. • Protists and Bacteria have, on average, larger environmental niche breadths. • On average, fungi proportionally harbor more generalist taxa while bacteria harbor more specialist taxa than any other group. • Small niche breadth towards environmental extremes suggests increased specialization. [ABSTRACT FROM AUTHOR]

Abstract: Little is known about the diversity and ecology of earthworms in floodplains, as well as their response to natural and anthropic perturbations (e.g. floods, river channelisation, floodplain restoration). We characterised the patterns of earthworm communities and species traits in the different habitats of a lowland restored floodplain in Switzerland. In addition to classical species-based metrics, such as species richness and Shannon diversity, species traits were used to calculate the community weighted means (CWMs) of traits and functional dispersion (FDis). We hypothesised that trait-based metrics would reveal clearer patterns than classical approaches. The distribution of earthworm traits varied among habitats in relation to changes in flooding frequency: poorly developed gravel bar soils most exposed to flooding were characterised by high abundance of small epigeic species and low abundance of large anecic species. Differences in anecic and endogeic earthworm community structure matched flood frequency. In agreement with our hypothesis, CWMs were more strongly correlated to environmental variables than species composition, diversity, or functional diversity. Based on these results, the ratio of the relative abundances of epigeic and anecic species, and the differences in species composition within anecic and endogeic ecological types of earthworms were identified as indicators of soil development in floodplains. [Copyright &y& Elsevier]

Abstract: Among the non-pollen micro-fossils commonly encountered in Quaternary sediment samples prepared for pollen analysis are many shells of testate amoebae. Testate amoebae are eukaryotic micro-organisms which are increasingly used in ecological and palaeoecological studies, particularly as indicators of hydrological change in Sphagnum-dominated peatlands. In this study we address the extent to which testate amoebae are used in palynological research, the key challenges to more widespread use, and the extent to which ecological information is retained in the testate amoeba assemblages of standard palynological slides. To achieve this we review the literature on the use of testate amoebae in palynology, compare testate amoeba records produced by palynological and water-based preparation methods and carry out simulations using previously-derived datasets. Our results show that testate amoebae are widely encountered in Quaternary palynological studies, primarily in peatlands, but the information which they can provide is undermined by limited taxonomic knowledge. Many taxa are destroyed in pollen preparations, but for taxa that are retained patterns of abundance parallel those determined using water-based preparation methods. Although the loss of sensitive taxa limits the ecological information contained in testate amoeba assemblages the information preserved is likely to be useful in a multiproxy approach to palaeoenvironmental reconstruction. To help improve taxonomic awareness and encourage the use of testate amoebae in palynology we present a basic introduction to testate amoeba taxonomy and a guide to the taxonomic literature. [Copyright &y& Elsevier]

Abstract: Functional traits (FT) offer a new framework to understand the ecology of organisms and overcome taxonomic difficulties that currently limit the study of minute soil taxa. FT are likely to be selected by environmental filters and hence they may provide more direct information on ecosystem characteristics than the species composition of a community. We tested the potential of testate amoeba (TA) functional traits as bioindicators of selected ecosystem processes in the context of a restored floodplain in north-western Switzerland. The floodplain was divided into six functional process zones (FPZs) associated to distinct post-restoration successional stages. We selected TA FT and computed three functional indices: functional richness (FRic), divergence (FDiv), evenness (FEve), and dispersion (FDis). We then compared the patterns of functional indices and classical diversity indices such as species richness, diversity and evenness. We assessed whether traits converged or were over-dispersed in the different FPZs using a randomization procedure. Finally, we related environmental variables and functional traits using the “Fourth Corner” statistic. This procedure enabled us to highlight relations that can potentially be used for bioindication. Promising candidates include the relationships between shell biovolume and vegetation structure and between shell compression and plant litter input variables. [Copyright &y& Elsevier]

Abstract: Peatlands represent massive global C pools and sinks. Carbon accumulation depends on the ratio between net primary production and decomposition, both of which can change under projected increases of atmospheric CO2 and N deposition. The decomposition of litter is influenced by 1) the quality of the litter, and 2) the microenvironmental conditions in which the litter decomposes. This study aims at experimentally testing the effects of these two drivers in the context of global change. We studied the in situ litter decomposition from three common peatland species (Eriophorum vaginatum, Polytrichum strictum and Sphagnum fallax) collected after one year of litter production under pre-treatment conditions (elevated CO2: 560 ppm or enhanced N: 3 g m−2 y−1 NH4NO3) and decomposed the following year under treatment conditions (same as pre-treatment). By considering the cross-effects between pre-treatments and treatments, we distinguished between the effects on mass loss of 1) the pre-treatment-induced litter quality and 2) the treatment conditions under which the litters were decomposing. The combination between CO2 pre-treatment and CO2 treatment reduced Polytrichum decomposition by −24% and this can be explained by litter quality-driven decomposition changes brought by the pre-treatment. CO2 pre-treatment reduced Eriophorum litter quality, although this was not sufficient to predict decomposition. The N addition pre-treatment reduced the decomposition of Eriophorum, due to enhanced lignin and soluble phenols concentrations in the initial litter, and reduced litter-driven losses of starch and enhanced litter-driven losses of soluble phenols. While decomposition indices based on initial litter quality provide a broad explanation of quantitative and qualitative decomposition, they can only be taken as first approximations. Indeed, the microbial ATP activity, the litter N loss and resulting litter quality, were strongly altered irrespective of the compounds'' initial concentration and by means of processes that occurred independently of the initial litter-qualitative changes. The experimental design was valuable to assess litter- and ecosystem-driven decomposition pathways simultaneously or independently. The ability to separate these two drivers makes it possible to attest the presence of litter-qualitative changes even without any litter biochemical determinations, and shows the screening potential of this approach for future experiments dealing with multiple plant species. [Copyright &y& Elsevier]

Testate amoebae (TA) are a common and diverse group of protists and are especially abundant in peatlands. The structure of peatland TA communities is well correlated to surface moisture and water table depth (WTD). For that reason, TA are widely used as proxy indicators in ecological and palaeoecological studies. Peatlands are abundant across Asia, but the diversity and ecology of the TA that inhabit these systems are poorly documented. It is therefore unclear whether TA can be used as palaeohydrological indicators in the manner in which they commonly are in Europe and North-America. There is particular uncertainty as to the efficacy of this approach in the lower latitudes. We compiled existing and new data on testate amoebae from 1124 Sphagnum -dominated samples from 42 individual peatlands covering broad latitudinal (25°–66° N) and longitudinal (68°–129° E) ranges. Using a consensus taxonomic framework, we built a checklist of TA and developed TA-based hydrological transfer functions for Asian peatlands. The results showed that three models, weighted averaging (WA), weighted average partial least squares (WA-PLS), and maximum likelihood (ML), predicted similar WTD values for full samples, while the modern analogue technique (MAT) produced the strongest (R2 boot = 0.58) relationship between observed and estimated water-table depths (WTDs). Removing outlier samples improved the R2 values of observed vs. estimated WTDs, with ML then demonstrating the strongest predictive power (R2 boot = 0.68, RMSEP boot = 8.98 cm). The predictive capability of the developed WTD transfer function is comparable to equivalent models for Europe and North America and thus can be used for palaeohydrological reconstructions for boreal to subtropical peatlands in Asia. • A testate amoebae taxonomic checklist was compiled from Asian peatlands. • We developed continental scale testate amoeba-based hydrological transfer functions. • The water-table depth transfer functions can be used for paleohydrological reconstructions. [ABSTRACT FROM AUTHOR]

• We compared the diversity of protists in freshwater, marine plankton and soil. • Soil hosts the highest protist γ diversity, followed by oceans and freshwater. • Phagocytosis seems to be the dominant process in eukaryotic nutrient acquisition. • Relative abundance of phototrophs in soils is comparable to marine systems. Protists dominate eukaryotic diversity and play key functional roles in all ecosystems, particularly by catalyzing carbon and nutrient cycling. To date, however, a comparative analysis of their taxonomic and functional diversity that compares the major ecosystems on Earth (soil, freshwater and marine systems) is missing. Here, we present a comparison of protist diversity based on standardized high throughput 18S rRNA gene sequencing of soil, freshwater and marine environmental DNA. Soil and freshwater protist communities were more similar to each other than to marine protist communities, with virtually no overlap of Operational Taxonomic Units (OTUs) between terrestrial and marine habitats. Soil protists showed higher γ diversity than aquatic samples. Differences in taxonomic composition of the communities led to changes in a functional diversity among ecosystems, as expressed in relative abundance of consumers, phototrophs and parasites. Phototrophs (eukaryotic algae) dominated freshwater systems (49% of the sequences) and consumers soil and marine ecosystems (59% and 48%, respectively). The individual functional groups were composed of ecosystem- specific taxonomic groups. Parasites were equally common in all ecosystems, yet, terrestrial systems hosted more OTUs assigned to parasites of macro-organisms while aquatic systems contained mostly microbial parasitoids. Together, we show biogeographic patterns of protist diversity across major ecosystems on Earth, preparing the way for more focused studies that will help understanding the multiple roles of protists in the biosphere. [ABSTRACT FROM AUTHOR]

The use of synthetic pesticides in agriculture is increasingly debated. However, few studies have compared the impact of synthetic pesticides and alternative biopesticides on non-target soil microorganisms playing a central role in soil functioning. We conducted a mesocosm experiment and used high-throughput amplicon sequencing to test the impact of a fungal biopesticide and a synthetic fungicide on the diversity, the taxonomic and functional compositions, and co-occurrence patterns of soil bacterial, fungal and protist communities. Neither the synthetic pesticide nor the biopesticide had a significant effect on microbial α-diversity. However, both types of pesticides decreased the complexity of the soil microbial network. The two pesticides had contrasting impacts on the composition of microbial communities and the identity of key taxa as revealed by microbial network analyses. The biopesticide impacted keystone taxa that structured the soil microbial network. The synthetic pesticide modified biotic interactions favouring taxa that are less efficient at degrading organic compounds. This suggests that the biopesticides and the synthetic pesticide have different impact on soil functioning. Altogether, our study shows that pest management products may have functionally significant impacts on the soil microbiome even if microbial α-diversity is unaffected. It also illustrates the potential of high-throughput sequencing analyses to improve the ecotoxicological risk assessment of pesticides on non-target soil microorganisms. Unlabelled Image • The impact of a synthetic pesticide and a biopesticide on soil microbes was tested. • Impact on microbial α-diversity was weak, transient and compound-specific. • Both pesticides altered co-occurrences of microbial taxa and community composition. • Both the biopesticide and the synthetic pesticide can affect soil microorganisms. • Measures of α-diversity are insufficient to evaluate the impact of pesticide use. [ABSTRACT FROM AUTHOR]

Beta diversity is a key component of biodiversity with implications ranging from species dynamics to ecosystem functioning. However, β-diversity and its drivers have received little attention, especially for micro-eukaryotes which play key roles in soil functioning. We studied the diversity of soil micro-eukaryotes in a Swiss lowland floodplain using high-throughput Illumina sequencing of soil DNA. We determined the temporal vs. spatial patterns of soil micro-eukaryotic α- and β-diversity in six contrasted habitats sampled over one year. We identified the drivers of these patterns among soil conditions and functions and identified indicator taxa of habitats in each season. We found higher spatial than temporal variability and a strong space-time interaction in soil micro-eukaryotic diversity patterns as well as in their edaphic drivers, which contrasts with previous observation of bacterial diversity patterns. Our results show that, although soil micro-eukaryotic diversity indeed varies seasonally, it is correlated most strongly with edaphic variables and vegetation but the strength of correlations with individual drivers varied seasonally. Microbial diversity patterns and their drivers can thus differ quite substantially among seasons and taxa. Despite the dominance of spatial patterns, the temporal component of microbial diversity should not be ignored to accurately estimate the diversity and the complexity of soil microbial community assembly processes. Given the importance of soil microbial diversity for ecosystem functioning such knowledge is relevant for land management. Image 1 • We studied floodplain soil micro-eukaryotic diversity by Illumina sequencing of e-DNA. • Alpha and beta diversity varied more spatially among habitats than seasonally. • The edaphic drivers of α and β diversities changed among habitats and seasons. • These patterns reflect seasonal flood dynamics and soil biogeochemical cycles. • Thus, microbial ecology of dynamic systems should best be studied over time and space. [ABSTRACT FROM AUTHOR]

A high diversity of Apicomplexa was recently found in tropical soils presumably reflecting the diversity of their invertebrate hosts, but such patterns have not been explored in colder regions. We analysed the diversity of Apicomplexa and their potential metazoan hosts in litter and mosses collected in 11 different alpine habitats using an eDNA metabarcoding approach. The abundance and diversity of Apicomplexa phylotypes and of their potential invertebrate hosts were positively correlated. This confirms that eDNA metabarcoding is a useful tool to explore the unknown biodiversity of free-living eukaryotes, as well as potential host-parasite interactions. Future studies should aim at describing this diversity using a combination of morphological and molecular approaches. • We found a high diversity of Apicomplexa sequences in alpine moss and litter eDNA. • Read numbers of Apicomplexa and their Metazoa hosts are positively correlated. • EDNA metabarcoding allows a fast comparison of Apicomplexa and Metazoa diversity. [ABSTRACT FROM AUTHOR]

Summary Life was microbial for the majority of Earth's history, but as very few microbial lineages leave a fossil record, the Precambrian evolution of life remains shrouded in mystery. Shelled (testate) amoebae stand out as an exception with rich documented diversity in the Neoproterozoic as vase-shaped microfossils (VSMs). While there is general consensus that most of these can be attributed to the Arcellinida lineage in Amoebozoa, it is still unclear whether they can be used as key fossils for interpretation of early eukaryotic evolution. Here, we present a well-resolved phylogenomic reconstruction based on 250 genes, obtained using single-cell transcriptomic techniques from a representative selection of 19 Arcellinid testate amoeba taxa. The robust phylogenetic framework enables deeper interpretations of evolution in this lineage and demanded an updated classification of the group. Additionally, we performed reconstruction of ancestral morphologies, yielding hypothetical ancestors remarkably similar to existing Neoproterozoic VSMs. We demonstrate that major lineages of testate amoebae were already diversified before the Sturtian glaciation (720 mya), supporting the hypothesis that massive eukaryotic diversification took place in the early Neoproterozoic and congruent with the interpretation that VSM are arcellinid testate amoebae. Highlights • Arcellinida testate amoebae are comprised of seven major lineages • Reconstructed hypothetical ancestral states are congruent with Tonian fossils • Combined analysis of phylogenies and fossils suggest divergence as early as 730 mya Arcellinid testate amoebae are some of the earliest eukaryotic microbes. Lahr et al. show that these amoebae have seven major deep phylogenetic lineages, probably diverged as early as 730 mya. Diversification of these microbes in the mid-Neoproterozoic was probably happening alongside the gradual oxygenation of oceans. [ABSTRACT FROM AUTHOR]