Peatland Testate Amoebae Research Articles

Hyalosphenia papilio paynei subsp. nov. – a highly conspicuous and localized Sphagnum peatland testate amoeba

Hyalosphenia papilio paynei subsp. nov. – a highly conspicuous and localized Sphagnum peatland testate amoeba

Ecology of peatland testate amoebae in Svalbard and the development of transfer functions for reconstructing past water-table depth and pH

Peatlands are valuable archives of information about past environmental conditions and represent a globally-important carbon store. Robust proxy methods are required to reconstruct past ecohydrological dynamics in high-latitude peatlands to improve our understanding of change in these carbon-rich ecosystems. The High Arctic peatlands in Svalbard are at the northern limit of current peatland distribution and have experienced rapidly rising temperatures of 0.81 °C per decade since 1958. We examine the ecology of peatland testate amoebae in surface vegetation samples from permafrost peatlands on Spitsbergen, the largest island of the Svalbard archipelago, and develop new transfer functions to reconstruct water-table depth (WTD) and pH that can be applied to understand past peatland ecosystem dynamics in response to climate change. These transfer functions are the first of their kind for peatlands in Svalbard and the northernmost developed to date. Multivariate statistical analysis shows that WTD and pore water pH are the dominant controls on testate amoeba species distribution. This finding is consistent with results from peatlands in lower latitudes with regard to WTD and supports work showing that when samples are taken across a long enough trophic gradient, peatland trophic status is an important control on the distribution of testate amoebae. No differences were found between transfer functions including and excluding the taxa with weak idiosomic tests (WISTs) that are most susceptible to decay. The final models for application to fossil samples therefore excluded these taxa. The WTD transfer function demonstrates the best performance (R2LOO = 0.719, RMSEPLOO = 3.2 cm), but the pH transfer function also performs well (R2LOO = 0.690, RMSEPLOO = 0.320). The transfer functions were applied to a core from western Spitsbergen and suggest drying conditions ~1750 CE, followed by a trend of recent wetting and increasing pH from ~1920 CE. These new transfer functions allow the reconstruction of past peatland WTD and pH in Svalbard, thereby enabling a greater understanding of long-term ecohydrological dynamics in these rapidly changing ecosystems.

Recent Changes in Peatland Testate Amoeba Functional Traits and Hydrology Within a Replicated Site Network in Northwestern Québec, Canada

Northern peatlands, which are highly heterogeneous ecosystems, are a globally important carbon (C) store. Understanding the drivers and predicting the future trajectory of the peatland C store requires upscaling from cores and sites to regions and continents, alongside a detailed understanding of the mechanisms governing their C sequestration. Studies incorporating replication are therefore important to quantify how peatland heterogeneity may affect upscaling from local-scale dynamics to models. In addition, we need to better understand the processes driving observed variability, but the interplay between plants, microbes and C cycling in peatlands remains poorly understood. One approach to address both issues is to examine replicated microbiological functional traits within a multi-proxy framework to provide an ecosystem-level perspective on ecological and biogeochemical processes. Peatland testate amoebae are a functionally important group of protists that are well suited to such an approach. Analysing testate amoeba functional traits provides an opportunity to examine processes that may affect key peatland ecosystem services, such as C sequestration. Here, we compared four key testate amoeba functional traits (mixotrophy, biovolume, aperture size and aperture position) to C accumulation, hydrological and vegetation changes in 12 post-Little Ice Age peat records. Samples were collected from high-boreal and low-subarctic regions in northwestern Quebec, Canada in an experimental design that includes internal and external replication at both site and regional scales. Our results showed that correspondence between C accumulation, hydrology and testate amoeba functional traits varied, but recent changes in mixotrophy and aperture size, which may affect peatland C sequestration potential and microbial food web structure respectively, showed tentative links to recent C accumulation increases. Vegetation, especially Sphagnum abundance was important in promoting mixotrophy and small aperture size in testate amoeba communities. Future impacts of climate change on peatland vegetation will further influence the functional role of testate amoebae on C sequestration through changing mixotrophic testate amoeba abundance.

Ecology and paleoenvironmental application of testate amoebae in peatlands of the high-elevation Colombian páramo

AbstractWe investigated the ecology and paleoecology of testate amoebae in peatlands of the Colombian páramo to assess the use of testate amoebae as paleoenvironmental indicators. Objectives were to (1) identify environmental controls on testate amoebae, (2) develop transfer functions for paleoenvironmental inference, and (3) examine testate amoebae in a Holocene peat core and compare our findings with other proxy records. Results from 96 modern samples indicate that testate amoebae are sensitive to pH and surface moisture, and cross-validation of transfer functions indicates potential for paleoenvironmental applications. Testate amoebae from the Triunfo Peatland in the Central Cordillera provided a proxy record of pH and water-table depth for the late Holocene, and inferred changes were correlated with peat C/N measurements during most of the record. Comparison with a lake-level reconstruction suggests that at least the major testate amoeba–inferred changes were driven by climate. Our work indicates that testate amoebae are useful paleoenvironmental indicators in high-elevation tropical peatlands.

Towards a Holarctic synthesis of peatland testate amoeba ecology: Development of a new continental-scale palaeohydrological transfer function for North America and comparison to European data

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.

Ecology of peatland testate amoebae in the Alaskan continuous permafrost zone

Arctic peatlands represent a major global carbon store, but rapid warming poses a threat to their long-term stability. Testate amoebae are sensitive hydrological indicators that offer insight into Holocene environmental change in peatlands. However, in contrast to temperate peatlands, there have only been a few studies into the ecology of testate amoebae and their efficacy as environmental indicators in permafrost peatlands. We present the first study of testate amoeba ecology from peatlands in the continuous permafrost zone, based on samples from across the Alaskan North Slope. Multivariate statistical analyses show that pore water electrical conductivity (EC), a proxy for nutrient status along the ombrotrophic-minerotrophic gradient, is the dominant control on testate amoeba distribution. Water-table depth (WTD) is also a significant control on testate amoeba distribution, but is secondary to EC. We present two new testate amoeba-based transfer functions to reconstruct both EC (TFEC) and WTD (TFWTD), the first for peatlands in the continuous permafrost zone. The transfer functions are based on Weighted Averaging Partial Least Squares (WAPLS) regression and were assessed using leave-one-out (LOO) cross-validation. We find that both transfer functions have good predictive power. TFWTD is the best performing model (R2JACK = 0.84, RMSEPJACK = 6.66 cm), but TFEC also performs well (R2JACK = 0.76, RMSEPJACK = 146 μS cm−1). Our findings are similar to those conducted in peatlands in discontinuous permafrost regions. The new transfer functions open the opportunity for reconstructing the Holocene dynamics of peatlands of the continuous permafrost zone in Alaska, which represent rapidly changing ecosystems.

Taxonomic and functional traits responses of Sphagnum peatland testate amoebae to experimentally manipulated water table

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: −4cm, intermediate: −15cm, and dry: −25cm) 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.

What is the optimum sample size for the study of peatland testate amoeba assemblages?

Testate amoebae are widely used in ecological and palaeoecological studies of peatlands, particularly as indicators of surface wetness. To ensure data are robust and comparable it is important to consider methodological factors which may affect results. One significant question which has not been directly addressed in previous studies is how sample size (expressed here as number of Sphagnum stems) affects data quality. In three contrasting locations in a Russian peatland we extracted samples of differing size, analysed testate amoebae and calculated a number of widely-used indices: species richness, Simpson diversity, compositional dissimilarity from the largest sample and transfer function predictions of water table depth. We found that there was a trend for larger samples to contain more species across the range of commonly-used sample sizes in ecological studies. Smaller samples sometimes failed to produce counts of testate amoebae often considered minimally adequate. It seems likely that analyses based on samples of different sizes may not produce consistent data. Decisions about sample size need to reflect trade-offs between logistics, data quality, spatial resolution and the disturbance involved in sample extraction. For most common ecological applications we suggest that samples of more than eight Sphagnum stems are likely to be desirable.

Contrasting diversity of testate amoebae communities in Sphagnum and brown-moss dominated patches in relation to shell counts

Ecological studies of peatland testate amoebae are generally based on totals of 150 individuals per sample. However, the suitability of this standard has never been assessed for alkaline habitats such as spring fens. We explored the differences in testate amoeba diversity between Sphagnum and brown-moss microhabitats at a mire site with a highly diversified moss layer which reflects the small-scale heterogeneity in groundwater chemistry. Relationships between sampling efficiency and sample completeness were explored using individual-based species accumulation curves and the effort required to gain an extra species was assessed. Testate amoeba diversity differed substantially between microhabitats, with brown mosses hosting on average twice as many species and requiring greater shell totals to reach comparable sample analysis efficiency as for Sphagnum. Thus, for samples from alkaline conditions an increase in shell totals would be required and even an overall doubling up to 300 individuals might be considered for reliable community description. Our small-scale data are likely not robust enough to provide an ultimate solution for the optimization of shell totals. However, the results proved that testate amoebae communities from acidic and alkaline environments differ sharply in both species richness and composition and they might call for different methodological approaches.

Exploring the diversity and ecology of testate amoebae in West Siberian peatlands

Testate amoebae are valued for their functional significance and application as indicators of environmental conditions, particularly in peatland ecosystems. Research on testate amoebae has increased dramatically in recent years but there are still large parts of the world which have seen very little research. Here we consider testate amoeba communities of the West Siberian Lowland, the world’s largest peatland region and therefore one of the largest potential habitats for testate amoebae. Extensive sampling identified 89 taxa and showed that testate amoeba communities are structured by their physical and biological environment. We identified significant relationships between amoeba communities and both moisture content and vegetation composition. Despite the assemblages containing many widely-distributed species, some taxa considered typical of peatlands (e.g. Archerella flavum and Hyalosphenia papilio) were comparatively rare or absent, paralleling findings further south in Asia. We suggest that testate amoebae in this region deserve further study and may have useful applications in palaeoecological reconstruction and as bioindicators of the impacts of oil and gas extraction.

Loss of testate amoeba functional diversity with increasing frost intensity across a continental gradient reduces microbial activity in peatlands

Soil microbial communities significantly contribute to global fluxes of nutrients and carbon. Their response to climate change, including winter warming, is expected to modify these processes through direct effects on microbial functions due to osmotic stress, and changing temperature regimes. Using four European peatlands reflecting different frequencies of frost events, we show that peatland testate amoeba communities diverge among sites with different winter climates, and that this is reflected through contrasting functions. We found that exposure to harder soil frost promoted species β-diversity (species turnover) thus shifting the community composition of testate amoebae. In particular, we found that harder soil frost, and lower water-soluble phenolic compounds, induced functional turnover through the decrease of large species (−68%, >80μm) and the increase of small-bodied mixotrophic species (i.e. Archerella flavum; +79%). These results suggest that increased exposure to soil frost could be highly limiting for large species while smaller species are more resistant. Furthermore, we found that β-glucosidase enzymatic activity, in addition to soil temperature, strongly depended of the functional diversity of testate amoebae (R2=0.95, ANOVA). Changing winter conditions can therefore strongly impact peatland decomposition process, though it remains unclear if these changes are carried-over to the growing season.

Tree encroachment may lead to functionally-significant changes in peatland testate amoeba communities

Climate change is likely to cause increased tree recruitment on open peatlands but we currently have little idea what consequences this vegetation change may have below-ground. Here we use transects across forested to open bog ecotones at three Russian peatland complexes to assess potential changes in the most abundant group of peatland protists – the testate amoebae. We show that the testate amoeba communities of forested and open bog are markedly different with a very abrupt boundary at, or near, the vegetation ecotone. Changes along our transects suggest that tree encroachment may reduce the trophic level of testate amoeba communities and reduce the contribution of mixotrophic testate amoebae to primary production. Our study strongly suggests that increased tree recruitment on open peatlands will have important consequences for both microbial biodiversity and microbially-mediated ecosystem processes.

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.

Long-term hydrological dynamics and fire history over the last 2000 years in CE Europe reconstructed from a high-resolution peat archive

Sphagnum peatlands in the oceanic-continental transition zone of Poland are currently influenced by climatic and anthropogenic factors that lead to peat desiccation and susceptibility to fire. Little is known about the response of Sphagnum peatland testate amoebae (TA) to the combined effects of drought and fire. To understand the relationships between hydrology and fire dynamics, we used high-resolution multi-proxy palaeoecological data to reconstruct 2000 years of mire history in northern Poland. We employed a new approach for Polish peatlands – joint TA-based water table depth and charcoal-inferred fire activity reconstructions. In addition, the response of most abundant TA hydrological indicators to charcoal-inferred fire activity was assessed. The results show four hydrological stages of peatland development: moderately wet (from ∼35 BC to 800 AD), wet (from ∼800 to 1390 AD), dry (from ∼1390 to 1700 AD) and with an instable water table (from ∼1700 to 2012 AD). Fire activity has increased in the last millennium after constant human presence in the mire surroundings. Higher fire activity caused a rise in the water table, but later an abrupt drought appeared at the onset of the Little Ice Age. This dry phase is characterized by high ash contents and high charcoal-inferred fire activity. Fires preceded hydrological change and the response of TA to fire was indirect. Peatland drying and hydrological instability was connected with TA community changes from wet (dominance of Archerella flavum, Hyalosphenia papilio, Amphitrema wrightianum) to dry (dominance of Cryptodifflugia oviformis, Euglypha rotunda); however, no clear fire indicator species was found. Anthropogenic activities can increase peat fires and cause substantial hydrology changes. Our data suggest that increased human fire activity was one of the main factors that influenced peatland hydrology, though the mire response through hydrological changes towards drier conditions was delayed in relation to the surrounding vegetation changes.

Evaluating the use of testate amoebae for palaeohydrological reconstruction in permafrost peatlands

The melting of high-latitude permafrost peatlands is a major concern due to a potential positive feedback on global climate change. We examine the ecology of testate amoebae in permafrost peatlands, based on sites in Sweden (~ 200 km north of the Arctic Circle). Multivariate statistical analysis confirms that water-table depth and moisture content are the dominant controls on the distribution of testate amoebae, corroborating the results from studies in mid-latitude peatlands. We present a new testate amoeba-based water table transfer function and thoroughly test it for the effects of spatial autocorrelation, clustered sampling design and uneven sampling gradients. We find that the transfer function has good predictive power; the best-performing model is based on tolerance-downweighted weighted averaging with inverse deshrinking (performance statistics with leave-one-out cross validation: R2 = 0.87, RMSEP = 5.25 cm). The new transfer function was applied to a short core from Stordalen mire, and reveals a major shift in peatland ecohydrology coincident with the onset of the Little Ice Age (c. AD 1400). We also applied the model to an independent contemporary dataset from Stordalen and find that it outperforms predictions based on other published transfer functions. The new transfer function will enable palaeohydrological reconstruction from permafrost peatlands in Northern Europe, thereby permitting greatly improved understanding of the long-term ecohydrological dynamics of these important carbon stores as well as their responses to recent climate change.

Eight species in the Nebela collaris complex: Nebela gimlii (Arcellinida, Hyalospheniidae), a new species described from a Swiss raised bog

We describe here a new species of sphagnicolous testate amoeba found abundantly in the forested part of the Le Cachot peatland (Jura Mountains, Neuchâtel, Switzerland) based on microscopical observations (LM, SEM). The new species, called Nebela gimlii was placed in a phylogenetic tree based on mitochondrial cytochrome oxidase sequences (COI), and branched robustly within the N. collaris complex next to the morphologically similar N. guttata and N. tincta. It is however genetically clearly distinct from these two species, and differs morphologically from them by its smaller size and stouter shape of the shell. This new species completes the phylogeny of the Nebela collaris species complex, with now eight species described, mostly from peatlands and acidic forest litter, and further demonstrates the existence of an unknown diversity within testate amoebae. Improving the taxonomy of testate amoebae in peatlands and clarifying the ecology of newly discovered species should make these organisms even more valuable as bioindicator and for palaeoecological reconstruction.

Seasonal changes in Sphagnum peatland testate amoeba communities along a hydrological gradient

Testate amoebae are an abundant and functionally important group of protists in peatlands, but little is known about the seasonal patterns of their communities. We investigated the relationships between testate amoeba diversity and community structure and water table depth and light conditions (shading vs. insolation) in a Sphagnum peatland in Northern Poland (Linje mire) in spring and summer 2010. We monitored the water table at five sites across the peatland and collected Sphagnum samples in lawn and hummock micro-sites around each piezometer, in spring (3 May) and mid-summer (6 August) 2010. Water table differed significantly between micro-sites and seasons (Kruskal–Wallis test, p=0.001). The community structure of testate amoebae differed significantly between spring and summer in both hummock and lawn micro-sites. We recorded a small, but significant drop in Shannon diversity, between spring and summer (1.76 vs. 1.72). Strongest correlations were found between testate amoeba communities and water table lowering and light conditions. The relative abundance of mixotrophic species Hyalosphenia papilio, Archerella flavum and of Euglypha ciliata was higher in the summer.

Response of Sphagnum Peatland Testate Amoebae to a 1-Year Transplantation Experiment Along an Artificial Hydrological Gradient

Peatland testate amoebae (TA) are well-established bioindicators for depth to water table (DWT), but effects of hydrological changes on TA communities have never been tested experimentally. We tested this in a field experiment by placing Sphagnum carpets (15 cm diameter) collected in hummock, lawn and pool microsites (origin) at three local conditions (dry, moist and wet) using trenches dug in a peatland. One series of samples was seeded with microorganism extract from all microsites. TA community were analysed at T0: 8–2008, T1: 5–2009 and T2: 8–2009. We analysed the data using conditional inference trees, principal response curves (PRC) and DWT inferred from TA communities using a transfer function used for paleoecological reconstruction. Density declined from T0 to T1 and then increased sharply by T2. Species richness, Simpson diversity and Simpson evenness were lower at T2 than at T0 and T1. Seeded communities had higher species richness in pool samples at T0. Pool samples tended to have higher density, lower species richness, Simpson diversity and Simpson Evenness than hummock and/or lawn samples until T1. In the PRC, the effect of origin was significant at T0 and T1, but the effect faded away by T2. Seeding effect was strongest at T1 and lowest vanished by T2. Local condition effect was strong but not in line with the wetness gradient at T1 but started to reflect it by T2. Likewise, TA-inferred DWT started to match the experimental conditions by T2, but more so in hummock and lawn samples than in pool samples. This study confirmed that TA responds to hydrological changes over a 1-year period. However, sensitivity of TA to hydrological fluctuations, and thus the accuracy of inferred DWT changes, was habitat specific, pool TA communities being least responsive to environmental changes. Lawns and hummocks may be thus better suited than pools for paleoecological reconstructions. This, however, contrasts with the higher prediction error and species’ tolerance for DWT with increasing dryness observed in transfer function models.Electronic supplementary materialThe online version of this article (doi:10.1007/s00248-014-0367-8) contains supplementary material, which is available to authorized users.

Ecology of testate amoebae in peatlands of central China and development of a transfer function for paleohydrological reconstruction

Testate amoebae are a diverse and abundant group of protozoa that constitute a large proportion of biomass in many ecosystems and probably fill important roles in ecosystem function. These microorganisms have attracted the interest of paleoecologists because the preserved shells of testate amoebae and their known hydrological preferences enable reconstruction of past hydrological change. In ombrotrophic peatlands, surface wetness reflects hydroclimate, so testate amoebae can play an important role in reconstruction of Holocene climate change. Previous studies, however, have been geographically restricted, mostly to North America and Europe. We studied the ecology of testate amoebae in peatlands from central China in relation to hydrology, pH and metal concentrations. We found that testate amoeba community structure was correlated with depth to water table (DWT) and that the hydrological preferences of species generally matched those of previous studies. We developed a weighted average DWT transfer function that enables prediction of water table depth with a cross-validated mean error of <5 cm. Our results demonstrate the potential for using testate amoebae to reconstruct paleohydrology in China. Such studies could contribute to our understanding of Holocene climate changes in China, particularly regarding past Asian monsoon activity.

Can pollution bias peatland paleoclimate reconstruction?

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.