The Impact of the Species–Area Relationship on Estimates of Paleodiversity

Estimates of paleodiversity patterns through time have relied on datasets that lump taxonomic occurrences from geographic areas of varying size per interval of time. In essence, such estimates assume that the species–area effect, whereby more species are recorded from larger geographic areas, is negligible for fossil data. We tested this assumption by using the newly developed Miocene Mammal Mapping Project database of western North American fossil mammals and its associated analysis tools to empirically determine the geographic area that contributed to species diversity counts in successive temporal bins. The results indicate that a species–area effect markedly influences counts of fossil species, just as variable spatial sampling influences diversity counts on the modern landscape. Removing this bias suggests some traditionally recognized peaks in paleodiversity are just artifacts of the species–area effect while others stand out as meriting further attention. This discovery means that there is great potential for refining existing time-series estimates of paleodiversity, and for using species–area relationships to more reliably understand the magnitude and timing of such biotically important events as extinction, lineage diversification, and long-term trends in ecological structure.     

Diversity biases in terrestrial mammalian assemblages and quantifying the differences between museum collections and published accounts: A case study from the Miocene of Nevada

Understanding diversity through time in the fossil record has primarily relied on the raw count of species within a given time interval, or species richness. These estimates are often derived from published fossil data, and standardized for sample size or geographic area. However, most methods that standardize richness by sample size are sensitive to changes in evenness, which introduces a potential problem with relying on published records: published accounts could be more even than the museum collections from which they are drawn. We address this bias in the context of mammalian paleodiversity, comparing published and museum collections of the Hemphillian Thousand Creek fauna to those of the Barstovian Virgin Valley fauna. We rarified specimen data, both number of identified specimens (NISP) and minimum number of individuals (MNI), and presence/absence data to compare published and museum data within and between faunas. Within faunas, published numbers of specimens are more even than museum samples, but the difference for localities in Virgin Valley is not significant. Neither published nor museum numbers of specimens indicate a significant difference between faunas, but the diversity pattern is reversed between the two data sets. Presence/absence rarefactions show no differences between sources; here, published data adequately sample the underlying museum records. Specimen-based evenness is not accurate in the published sample, and therefore we suggest that future studies of diversity in terrestrial mammalian assemblages must assess unpublished collections. Additionally, NISP data for Thousand Creek are more even than MNI data, suggesting that relying solely on NISP for assessing species diversity can also be misleading. Because publication bias alters richness and evenness, diversity estimates using published data must be circumspect about data sources.     

Biogeography of curimatid fishes reveals multiple lowland–upland river transitions and differential diversification in the Neotropics (Teleostei,Curimatidae)

The Neotropics harbors a megadiverse ichthyofauna comprising over 6300 species with approximately 80% in just three taxonomic orders within the clade Characiphysi. This highly diverse group has evolved in tropical South America over tens to hundreds of millions of years influenced mostly by re‐arrangements of river drainages in lowland and upland systems. In this study, we investigate patterns of spatial diversification in Neotropical freshwater fishes in the family Curimatidae, a species‐rich clade of the order Characiformes. Specifically, we examined ancestral areas, dispersal events, and shifts in species richness using spatially explicit biogeographic and macroevolutionary models to determine whether lowlands–uplands serve as museums or cradles of diversification for curimatids. We used fossil information to estimate divergence times in BEAST, multiple time‐stratified models of geographic range evolution in BioGeoBEARS, and alternative models of geographic state‐dependent speciation and extinction in GeoHiSSE. Our results suggest that the most recent common ancestor of curimatids originated in the Late Cretaceous likely in lowland paleodrainages of northwestern South America. Dispersals from lowland to upland river basins of the Brazilian and Guiana shields occurred repeatedly across independently evolving lineages in the Cenozoic. Colonization of upland drainages was often coupled with increased rates of net diversification in species‐rich genera such as Cyphocharax and Steindachnerina. Our findings demonstrate that colonization of novel aquatic environments at higher elevations is associated with an increased rate of diversification, although this pattern is clade‐dependent and driven mostly by allopatric speciation. Curimatids reinforce an emerging perspective that Amazonian lowlands act as a museum by accumulating species along time, whereas the transitions to uplands stimulate higher net diversification rates and lineage diversification.     

Species accumulation over space and time in European Plio-Holocene mammals

The rate of increase in species number with sampled area is one issue of major interest in ecology. Species number increases with sampled time as well, though this kind of analysis is much rarer in literature. Species-area and species-time relationships have been recently integrated in a single model, which allows studying how time and area interact with each other in determining the cumulative increase in species richness. Here we studied species-area, species-time, and species-time-area relationships in Plio-Holocene large mammals of Western Eurasia, by using an extensive database including 184 species distributed in 685 fossil sites. We found that the increase of species number with time is much higher than with area. When sampling inequality of fossil localities in time and space is accounted for, time and area interact with each other in a negative, though non-linear fashion. The intense climatic changes that characterized the Plio-Holocene period apparently affected both species-area and species-time relationships in large mammals, by increasing the slope of the former during the Pliocene and middle Pleistocene, and of the latter during younger, climatically harsher, late Pleistocene times. This study emphasizes the importance of accounting for time and space in tracing paleodiversity curves.     

Monitoring large herbivore diversity at different scales: comparing direct and indirect methods

Monitoring of large herbivores is central to research and management activities in many protected areas. Monitoring programs were originally developed to estimate (trends in) population sizes of individual species. However, emphasis is shifting increasingly towards conservation of diversity and communities instead of individual species, as a growing literature shows the importance of herbivore diversity for ecosystem functioning. We argue that the design of monitoring programs has not yet been adapted well to this new conservation paradigm. Using large herbivore census data from Hluhluwe-iMfolozi Park, South Africa, we studied how monitoring methodology (observational counts vs. dung counts) and spatial scale interact in influencing estimates of large herbivore species richness and diversity. Dung counts resulted in higher herbivore species richness and diversity estimates than direct observational counts, especially at finer monitoring resolutions (grid cells smaller than 25 km²). At monitoring resolutions coarser than 25 km² both methods gave comparable diversity estimates. The methods also yielded different spatial diversity estimates, especially at finer resolutions. Grid cells with high diversity according to the dung count data did not necessarily have high diversity according to the observational counts, as shown by low correlation of grid cell values of both methods. We discuss these results in the light of estimates of the sampling effort of each method and, hence, suggest new monitoring designs that are more suitable for tracking temporal and spatial trends in large herbivore diversity and community composition.     

Relative contribution of vegetation types to regional biodiversity in Central Zagross forests of Iran

Managers in conservation biology are continually faced with the dilemma of needing to demonstrate which areas should receive conservation priority based on the diversity of species contained. Darenasab (Hashtadpahloo) forest catchment with dominated oak species in Mediterranean forests of Zagross in Iran was chosen as a case study. In order to estimate plant species richness in different vegetation types at landscape level, field method that sample both trees and herbs strata simultaneously (modified multi-scale Whittaker plots) was used to make species–area curves. Twenty-one modified multi-scale Whittaker plots (250 m² area) were located randomly in four vegetation types. Three species–area, species–log (area) and log (species)–log (area) curves models were constructed. The log (species)–log (area) model had the highest adjusted r² among others. Based on Jaccard’s coefficient, the pure oak vegetation type was the most heterogeneous (22% overlap), and the cushion plants vegetation type is the most homogeneous (29% overlap). The slope of species–area curves had the least range (0.05) and the slope of species–log (area) curves had the largest range (4.38). When the slopes of species–log (area) curves divided by mean Jaccard’s coefficient, the species–log (area) curves estimated values closest to those observed. The index of vegetation types contribution to regional diversity for determining conservation priority in off-reserve area, based on ranking of the observed mean species per plots, slopes of the species–log (area) curves, mean of Jaccard’s coefficient, mean unique species per plot, and the number of threatened species in each vegetation types were calculated. This composite index may provide a scientific method to rank vegetation types with high conservation value.     

Plant diversity in sacred forest fragments of Western Ghats: a comparative study of four life forms

The effect of fragmentation on different life forms within tropical forest plant communities is poorly understood. We studied the effect of degree of fragmentation and surrounding matrix on trees, lianas, shrubs and epiphytes in tropical forest fragments of Kodagu, Western Ghats, India. These fragments exist as sacred groves amidst a highly modified agricultural landscape, and have been preserved by the religious sentiments of local communities. Plants were sampled at two sites in continuous forests and 11 forest fragments. A total of 122 species of trees, 29 species of lianas, 60 species of shrubs and 66 species of epiphytes were recorded. Trees exhibited a significant species–area relationship (R ² = 0.74). Richness estimates after controlling for stem density (rarefaction) revealed that observed species–area relationship was not an artefact of passive sampling. Variation in species richness of the other three groups was explained by stem density and structural diversity. Linear distance from the reserve forest did not explain any variation in species richness. All life forms exhibited significant nested pattern. Trees were nested along the area gradient while nestedness in the other three groups showed evidence in support of habitat nestedness. The four life forms thus responded variably to degree of fragmentation. Our study revealed that 74% of the regional diversity for trees was contributed by diversity among plots, highlighting the importance of inter-patch habitat diversity in maintaining the total regional species pool. We conclude that trees alone cannot serve as good indicator for taking appropriate conservation measures to mitigate species loss resulting from habitat fragmentation.     

The origin and evolution of coral species richness in a marine biodiversity hotspot*

The Coral Triangle (CT) region of the Indo‐Pacific realm harbors an extraordinary number of species, with richness decreasing away from this biodiversity hotspot. Despite multiple competing hypotheses, the dynamics underlying this regional diversity pattern remain poorly understood. Here, we use a time‐calibrated evolutionary tree of living reef coral species, their current geographic ranges, and model‐based estimates of regional rates of speciation, extinction, and geographic range shifts to show that origination rates within the CT are lower than in surrounding regions, a result inconsistent with the long‐standing center of origin hypothesis. Furthermore, endemism of coral species in the CT is low, and the CT endemics are older than relatives found outside this region. Overall, our model results suggest that the high diversity of reef corals in the CT is largely due to range expansions into this region of species that evolved elsewhere. These findings strongly support the notion that geographic range shifts play a critical role in generating species diversity gradients. They also show that preserving the processes that gave rise to the striking diversity of corals in the CT requires protecting not just reefs within the hotspot, but also those in the surrounding areas.     

Presence of cushion plants increases community diversity in the high equatorial Andes

Cushion plants are a common growth form in the equatorial páramo vegetation and their surfaces are often colonized by other plants. This paper analyzes the effect of the cushion plants on the community diversity at 4650 m on the eastern slope of the Iliniza volcano in Ecuador. Ninety sample plots of 1 m² size were located in the study area and were divided into 25 subplots in which presence and abundance of plant species was recorded. The community diversity was expressed as species richness, Simpson's diversity index, and evenness. Correlation between the cushion species and the composition of the colonists was measured with the CCA ordination analysis, correlation between the cushion cover and community diversity was measured by means of correlation analysis. Randomized species–area curves were used to compare richness of plant communities with and without the cushions. A total of 32 species were found including five cushion plants. Most species preferred to grow on the cushion surface whereas only a few species were able to colonize open ground. Species richness and Simpson's index were significantly correlated to the cushion area but no correlation was found for evenness. The cushions were usually composed of more than one species which hampered the examination of the cushion–colonist specific relationships. Nevertheless, cushions of Azorella and Arenaria seemed to provide more favorable habitat for colonization than the other cushion species. Comparison with an earlier study made on Iliniza indicates that the presence of the cushions significantly increases the richness of the plant community.     

Severe extinction and rapid recovery of mammals across the Cretaceous–Palaeogene boundary,and the effects of rarity on patterns of extinction and recovery

The end‐Cretaceous mass extinction ranks among the most severe extinctions of all time; however, patterns of extinction and recovery remain incompletely understood. In particular, it is unclear how severe the extinction was, how rapid the recovery was and how sampling biases might affect our understanding of these processes. To better understand terrestrial extinction and recovery and how sampling influences these patterns, we collected data on the occurrence and abundance of fossil mammals to examine mammalian diversity across the K‐Pg boundary in North America. Our data show that the extinction was more severe and the recovery more rapid than previously thought. Extinction rates are markedly higher than previously estimated: of 59 species, four survived (93% species extinction, 86% of genera). Survival is correlated with geographic range size and abundance, with widespread, common species tending to survive. This creates a sampling artefact in which rare species are both more vulnerable to extinction and less likely to be recovered, such that the fossil record is inherently biased towards the survivors. The recovery was remarkably rapid. Within 300 000 years, local diversity recovered and regional diversity rose to twice Cretaceous levels, driven by increased endemicity; morphological disparity increased above levels observed in the Cretaceous. The speed of the recovery tends to be obscured by sampling effects; faunas show increased endemicity, such that a rapid, regional increase in diversity and disparity is not seen in geographically restricted studies. Sampling biases that operate against rare taxa appear to obscure the severity of extinction and the pace of recovery across the K‐Pg boundary, and similar biases may operate during other extinction events.     

In Search of Glacial Refuges of the Land Snail Orcula dolium (Pulmonata,Orculidae) - An Integrative Approach Using DNA Sequence and Fossil Data

Harboring a large number of endemic species, the Alps and the Western Carpathians are considered as major centers of biodiversity. Nonetheless, the general opinion until the turn of the millennium was that both Central European mountain regions did not provide suitable habitat during the Last Glacial Maximum, but were colonized later from southern refuges. However, recent molecular genetic studies provide new evidence for peripheral Alpine refuges. We studied the phylogeography of the calciphilous land snail O. dolium across its distribution in the Alps and the Western Carpathians to assess the amount of intraspecific differentiation and to detect potential glacial refuges. A partial sequence of the mitochondrial COI was analyzed in 373 specimens from 135 sampling sites, and for a subset of individuals, partial sequences of the mitochondrial 16S and the nuclear histone H3 and H4 were sequenced. A molecular clock analysis was combined with a reconstruction of the species’ geographic range history to estimate how its lineages spread in the course of time. In order to obtain further information on the species’ past distribution, we also screened its extensive Pleistocene fossil record. The reconstruction of geographic range history suggests that O. dolium is of Western Carpathian origin and diversified already around the Miocene-Pliocene boundary. The fossil record supports the species’ presence at more than 40 sites during the last glacial and earlier cold periods, most of them in the Western Carpathians and the Pannonian Basin. The populations of O. dolium display a high genetic diversity with maximum intraspecific p-distances of 18.4% (COI) and 14.4% (16S). The existence of various diverged clades suggests the survival in several geographically separated refuges. Moreover, the sequence patterns provide evidence of multiple migrations between the Alps and the Western Carpathians. The results indicate that the Southern Calcareous Alps were probably colonized only during the Holocene.     

Conservation genetics in hypersaline inland waters: mitochondrial diversity and phylogeography of an endangered Iberian beetle (Coleoptera: Hydraenidae)

Saline inland waters are globally threatened habitats harbouring many specialised endemic species, which often have restricted geographic ranges, and occur as highly isolated populations. We studied the genetic variation and phylogeography of Ochthebius glaber Montes and Soler, a rare and endangered water beetle endemic to hypersaline streams in the South and Southeast of the Iberian Peninsula. We used a 633 bp fragment of cytochrome oxidase subunit 1 gene to determine the genetic diversity and phylogeographic structure within this species, and interpret this in the light of the speciesȁ9 conservation requirements. Thirteen populations were sampled across the speciesȁ9 geographic range, and genetic diversity found to be very high, with 37 haplotypes across the 71 specimens examined (p-distance 0.2–7.3%, average 3.1±0.4). Phylogeographic analyses revealed a surprisingly high degree of geographical structure, detectable among populations separated by relatively short geographical distances, with three main groups of haplotypes which have apparently been isolated for significant periods of time. Past fragmentation and contiguous range expansion events were inferred as the main causes of the detected geographical associations of haplotypes. The establishment of independent evolutionary lineages as conservation units is particularly important for species inhabiting saline habitats such as O. glaber, which is endangered by habitat loss across most of its distribution. However, given the natural instability of hypersaline environments, the conservation of a network of populations and potential habitats would be necessary to enable the preservation of the process generating and maintaining the diversity of the species.     

Anostracan cysts found in California salt lakes

The salt lakes of California are home to four genera and eight species of Anostraca. The cysts of most of these species are readily distinguished using a combination of cyst and geographic characters. Cyst identification is a valuable tool that can allow a look at fairy shrimp ecology when adults are not available. For example, fossil anostracan cysts from 500-year-old Mono Lake sediments show more character variation than currently exists in extant Artemia monica, indicating a greater diversity of Anostraca at that time. Continuing desertification, both natural and man-made, in the Mono Lake area over the last 500 years is associated with decreased diversity in the Mono Lake anostracan fauna.     

Combining counts and incidence data: an efficient approach for estimating the log-normal species abundance distribution and diversity indices

Obtaining accurate estimates of diversity indices is difficult because the number of species encountered in a sample increases with sampling intensity. We introduce a novel method that requires that the presence of species in a sample to be assessed while the counts of the number of individuals per species are only required for just a small part of the sample. To account for species included as incidence data in the species abundance distribution, we modify the likelihood function of the classical Poisson log-normal distribution. Using simulated community assemblages, we contrast diversity estimates based on a community sample, a subsample randomly extracted from the community sample, and a mixture sample where incidence data are added to a subsample. We show that the mixture sampling approach provides more accurate estimates than the subsample and at little extra cost. Diversity indices estimated from a freshwater zooplankton community sampled using the mixture approach show the same pattern of results as the simulation study. Our method efficiently increases the accuracy of diversity estimates and comprehension of the left tail of the species abundance distribution. We show how to choose the scale of sample size needed for a compromise between information gained, accuracy of the estimates and cost expended when assessing biological diversity. The sample size estimates are obtained from key community characteristics, such as the expected number of species in the community, the expected number of individuals in a sample and the evenness of the community.     

Evidence for a time-integrated species-area effect on the latitudinal gradient in tree diversity

The greater area of tropical forest biomes has been proposed as a factor that drives the latitudinal gradient in species diversity by modulating speciation and extinction rates. But speciation and extinction are processes that operate over millions of years, so an adequate test of area's contribution to diversity patterns must take into consideration that biome areas have changed through time in response to climate. Here we correlate estimates of current tree species diversity with a composite parameter integrating area over geological time for each continent's tropical, temperate, and boreal biomes. We find significant positive correlations between current tree diversity and area-time for periods since the Eocene, Oligocene, and Miocene, which we take as evidence for a time-integrated species-area effect on current patterns of species richness across biomes. These results contribute to explanations for why most lineages have tropical origins and why tropical forests are more diverse than extratropical forests.     

Spatiotemporal changes in diatom ecological profiles: implications for biomonitoring

Diatom indices developed in certain geographic regions are frequently used elsewhere, despite the strong evidence that such metrics are less useful when applied in regions other than that where species–environment relationships were originally assessed, showing that species have particular autoecological requirements in different geographic areas. In this study, we define the ecological profiles for selected environmental variables in three common epilithic diatom species in the Duero Basin (NW Spain), comparing our results with data gathered from different geographic regions. We assess differences in autoecological parameters obtained from the northern and southern subbasins, and from different years, in order to observe whether significant small-scale, spatiotemporal changes exist. Our results show that there are variations in species’ autoecological parameters between different regions. Furthermore, there are significant changes comparing northern and southern Duero subbasins for certain species and physiochemical variables. Additionally, different autoecologies have been observed for certain diatoms and environmental factors comparing two different years. It can be concluded that freshwater diatom autoecologies can vary between different geographic areas. This implies that, in order to improve water-quality bioassessments, autoecology-based diatom metrics should be developed by quantifying species distributions along environmental gradients, using datasets representative of the areas or river types where the metrics will be applied.     

Patterns of spatial genetic structure and diversity at the onset of a rapid range expansion: colonisation of the UK by the small red-eyed damselfly <Emphasis Type="Italic">Erythromma viridulum</Emphasis>

Species’ geographic ranges may vary in size in response to a change in environmental conditions. The specific genetic consequences of range expansions are context dependent, largely depending upon the rate of colonisation as well as the origins and numbers of founders, and the time since colonisation. Like other “charismatic” taxa, such as birds and lepidopterans, the distributions of odonates (dragonflies and damselflies) are well-known through substantial monitoring programmes co-ordinated by various societies. The small red-eyed damselfly Erythromma viridulum (Odonata: Zygoptera) has undergone a substantial, northward range expansion in Europe in the last 30 years and has recently-colonised two distinct areas in the UK. We quantify the immediate genetic consequences of this rapid colonisation by genotyping more than 1,400 E. viridulum from 39 sites across the northwest margin of this species’ geographic range. Levels of genetic diversity and spatial structure are impacted by this species recent range expansion and non-equilibrium conditions that drive weak genetic divergence, even at regional spatial scales. Populations of E. viridulum become less diverse towards the edge of this species’ distribution, presumably as a consequence of colonisation through a series of founder events. Specifically, there is a significant reduction in genetic diversity in the smallest, most recent focus of colonisation in the UK; however, there are generally low levels of genetic diversity across this E. viridulum’s northern range margin. While most populations are generally poorly differentiated, E. viridulum nonetheless consists of two distinct lineages that broadly differentiate between eastern and western Europe. Genetic divergence between the two UK colonisation foci are indicative of distinct immigration events from separate sources; however a general lack of spatial structure prevents us from pinpointing the specific origins of these migrant damselflies.     

U-series dating and taphonomy of Quaternary vertebrates from Brazilian caves

The geochronology and taphonomy of internationally important fossil bearing cave deposits were studied, both in the semi-arid Northern Bahia area and the subtropical southeastern Lagoa Santa area of Brazil. Taphonomic analysis suggests that the processes responsible for bone accumulation in the Brazilian caves vary between sites, and taphonomic bias can therefore be significant in causing differences in faunal composition. In the Toca da Boa Vista caves the presence of single articulated skeletons, and the entrance-related distribution indicate that random penetration of animals is the main mechanism of fossil accumulation, a process that biases the assemblage to smaller species, and takes place over extended time periods. In nearby Toca dos Ossos cave transport by runoff in the cave river is predominant, and biases the fauna remains to larger more robust bones and species. Deposition probably also occurred only at times of enhanced runoff giving a more contemporaneous assemblage. Similar processes were responsible for emplacement of the copious fossil remains in the more humid Lagoa Santa area, where terrigenous fossil deposits are found intercalated by massive speleothem calcite layers. In this area runoff under a drier climate probably accounts for the sediment emplacement inside caves. In both areas the mode of emplacement implies bias in the fossil record, resulting in fossil assemblages that do not mirror surface faunas, limiting palaeoenvironmental reconstruction.

Mass spectrometric U-series analysis of speleothem calcite overlaying fossil remains gives minimum ages for fossil deposition. These ages confirm the previous view that many of the deposits derive from the late glacial, but also show that much older material (some > 350,000 yr) is also present. The habitat requirements of critical fossil species such as bats and monkeys strongly suggest that they derive from much wetter periods when forest cover was present in the currently semi-arid Northern Bahia area. Taphonomy exerts a major control on the diversity and mode of emplacement of cave fossil deposits in eastern Brazil and thus detailed sedimentological and hydrological studies coupled with a sound geochronological approach are essential in quantifying the relative importance of each taphonomic processes before faunal and palaeoecological interpretations can be attempted.     

云南被子植物菊类分支的系统发育多样性及其分布格局

全球气候变化与人为活动等因素导致的生物多样性丧失,引起了全球各界对生物多样性保护的高度关注。传统生物多样性保护主要对物种、特有种、受威胁物种的种类组成及其分布模式开展研究,忽视了进化历史在生物多样性保护中的作用。云南是全球生物多样性热点地区的交汇区,生物多样性的保护历来受到广泛关注,为了更好地探讨云南生物多样性的保护措施,该研究以云南被子植物菊类分支物种为研究对象,基于物种间的演化关系,结合其地理分布,从进化历史的角度探讨物种、特有种、受威胁物种的种类组成及系统发育组成的分布格局,并整合自然保护地的空间分布,识别生物多样性的重点保护区域。结果表明：云南被子植物菊类分支的物种、特有种及受威胁物种的物种密度与系统发育多样性均显著正相关;通过零模型分析发现,由南向北标准化系统发育多样性逐渐降低;云南南部、东南部、西北部是云南被子植物菊类分支的重点保护区域,加强这些区域的保护,将最大化地保护生物多样性的进化历史和进化潜能。由此可见,融合进化历史信息的植物多样性格局分析不仅有助于更加深入地理解植物多样性的形成与演变,也为生物多样性保护策略的制定提供更多的思路。     

Fish pathogens near the Arctic Circle: molecular,morphological and ecological evidence for unexpected diversity of Diplostomum (Digenea: diplostomidae) in Iceland

Host–parasite systems at high latitudes are promising model systems for detecting and predicting the impact of accelerated environmental change. A major challenge is the lack of baselines for the diversity and distribution of parasites in Arctic wildlife, especially in the freshwater environment. Here we present the first known estimates of the species diversity and host associations of Diplostomum spp. in sub-Arctic freshwater ecosystems of the Palaearctic. Our analyses integrating different analytical approaches, phylogenies based on mitochondrial and nuclear DNA, estimates of genetic divergence, character-based barcoding, morphological examination, precise detection of microhabitat specialisation and host use, led to the discovery of one described and five putative new species that complete their life-cycles within a fairly narrow geographic area in Iceland. This increases the species richness of Diplostomum in Iceland by 200% and raises the number of molecularly characterised species from the Palaearctic to 17 species. Our results suggest that the diversity of Diplostomum spp. is underestimated globally in the high latitude ecosystems and call for a cautionary approach to pathogen identification in developing the much needed baselines of pathogen diversity that may help detect effects of climate change in the freshwater environment of the sub-Arctic.