The complete mitochondrial genome of Talpa martinorum (Mammalia: Talpidae), a mole species endemic to Thrace: genome content and phylogenetic considerations.

The complete mitogenome sequence of Talpa martinorum, a recently described Balkan endemic mole, was assembled from next generation sequence data. The mitogenome is similar to that of the three other Talpa species sequenced to date, being 16,835 bp in length, and containing 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, an origin of L-strand replication, and a control region or D-loop. Compared to other Talpa mitogenomes sequenced to date, that of T. martinorum differs in the length of D-loop and stop codon usage. TAG and T-- are the stop codons for the ND1 and ATP8 genes, respectively, in T. martinorum, whilst TAA acts as a stop codon for both ND1 and ATP8 in the other three Talpa species sequenced. Phylogeny reconstructions based on Maximum Likelihood and Bayesian inference analyses yielded phylogenies with similar topologies, demonstrating that T. martinorum nests within the western lineage of the genus, being closely related to T. aquitania and T. occidentalis.

Plasticity of thermal performance curves in a narrow range endemic water beetle.

Thermal history can plastically alter the response of ectotherms to temperature, and thermal performance curves (TPCs) are powerful tools for exploring how organismal-level performance varies with temperature. Plasticity in TPCs may be favoured in thermally variable habitats, where it can result in fitness benefits. However, thermal physiology remains insufficiently studied for freshwater insects despite freshwater biodiversity being at great risk under global change. Here, we assess how acclimation at either summer or winter average temperatures changes TPCs for locomotion activity and metabolism in Enochrus jesusarribasi (Hydrophilidae), a water beetle endemic to shallow saline streams in SE Spain. This beetle is a bimodal gas exchanger and so we also assessed how aerial and aquatic gas exchange varied across temperatures for both acclimation treatments. Responses of locomotory TPCs to thermal acclimation were relatively weak, but high temperature acclimated beetles tended to exhibit higher maximum locomotor activity and reduced TPC breadth than those acclimated at lower temperature. High temperature acclimation increased the thermal sensitivity of metabolic rates, contrary to the response generally found in aquatic organisms. Higher metabolic rates upon high temperature acclimation were achieved by increasing aerial, rather than aquatic oxygen uptake. Such plastic respiratory behaviour likely contributed to enhanced locomotor performance at temperatures around the optimum and thermal plasticity could thus be an important component in the response of aquatic insects to climate change. However, high temperature acclimation appeared to be detrimental for locomotion in subsequent exposure at upper sublethal temperatures, suggesting that this narrow range endemic may be vulnerable to future climate warming. This study demonstrates that TPCs are context-specific, differing with performance metric as well as thermal history. Such context dependency must be considered when using TPCs to predict organismal responses to climate change.

Universal metabolic constraints shape the evolutionary ecology of diving in animals.

Diving as a lifestyle has evolved on multiple occasions when air-breathing terrestrial animals invaded the aquatic realm, and diving performance shapes the ecology and behaviour of all air-breathing aquatic taxa, from small insects to great whales. Using the largest dataset yet assembled, we show that maximum dive duration increases predictably with body mass in both ectotherms and endotherms. Compared to endotherms, ectotherms can remain submerged for longer, but the mass scaling relationship for dive duration is much steeper in endotherms than in ectotherms. These differences in diving allometry can be fully explained by inherent differences between the two groups in their metabolic rate and how metabolism scales with body mass and temperature. Therefore, we suggest that similar constraints on oxygen storage and usage have shaped the evolutionary ecology of diving in all air-breathing animals, irrespective of their evolutionary history and metabolic mode. The steeper scaling relationship between body mass and dive duration in endotherms not only helps explain why the largest extant vertebrate divers are endothermic rather than ectothermic, but also fits well with the emerging consensus that large extinct tetrapod divers (e.g. plesiosaurs, ichthyosaurs and mosasaurs) were endothermic.

Water Beetles as Models in Ecology and Evolution.

Beetles have colonized water many times during their history, with some of these events involving extensive evolutionary radiations and multiple transitions between land and water. With over 13,000 described species, they are one of the most diverse macroinvertebrate groups in most nonmarine aquatic habitats and occur on all continents except Antarctica. A combination of wide geographical and ecological range and relatively accessible taxonomy makes these insects an excellent model system for addressing a variety of questions in ecology and evolution. Work on water beetles has recently made important contributions to fields as diverse as DNA taxonomy, macroecology, historical biogeography, sexual selection, and conservation biology, as well as predicting organismal responses to global change. Aquatic beetles have some of the best resolved phylogenies of any comparably diverse insect group, and this, coupled with recent advances in taxonomic and ecological knowledge, is likely to drive an expansion of studies in the future.

Phylogenomics of the superfamily Dytiscoidea (Coleoptera: Adephaga) with an evaluation of phylogenetic conflict and systematic error.

The beetle superfamily Dytiscoidea, placed within the suborder Adephaga, comprises six families. The phylogenetic relationships of these families, whose species are aquatic, remain highly contentious. In particular the monophyly of the geographically disjunct Aspidytidae (China and South Africa) remains unclear. Here we use a phylogenomic approach to demonstrate that Aspidytidae are indeed monophyletic, as we inferred this phylogenetic relationship from analyzing nucleotide sequence data filtered for compositional heterogeneity and from analyzing amino-acid sequence data. Our analyses suggest that Aspidytidae are the sister group of Amphizoidae, although the support for this relationship is not unequivocal. A sister group relationship of Hygrobiidae to a clade comprising Amphizoidae, Aspidytidae, and Dytiscidae is supported by analyses in which model assumptions are violated the least. In general, we find that both concatenation and the applied coalescent method are sensitive to the effect of among-species compositional heterogeneity. Four-cluster likelihood-mapping suggests that despite the substantial size of the dataset and the use of advanced analytical methods, statistical support is weak for the inferred phylogenetic placement of Hygrobiidae. These results indicate that other kinds of data (e.g. genomic meta-characters) are possibly required to resolve the above-specified persisting phylogenetic uncertainties. Our study illustrates various data-driven confounding effects in phylogenetic reconstructions and highlights the need for careful monitoring of model violations prior to phylogenomic analysis.

The chicken or the egg? Adaptation to desiccation and salinity tolerance in a lineage of water beetles.

Transitions from fresh to saline habitats are restricted to a handful of insect lineages, as the colonization of saline waters requires specialized mechanisms to deal with osmotic stress. Previous studies have suggested that tolerance to salinity and desiccation could be mechanistically and evolutionarily linked, but the temporal sequence of these adaptations is not well established for individual lineages. We combined molecular, physiological and ecological data to explore the evolution of desiccation resistance, hyporegulation ability (i.e., the ability to osmoregulate in hyperosmotic media) and habitat transitions in the water beetle genus Enochrus subgenus Lumetus (Hydrophilidae). We tested whether enhanced desiccation resistance evolved before increases in hyporegulation ability or vice versa, or whether the two mechanisms evolved in parallel. The most recent ancestor of Lumetus was inferred to have high desiccation resistance and moderate hyporegulation ability. There were repeated shifts between habitats with differing levels of salinity in the radiation of the group, those to the most saline habitats generally occurring more rapidly than those to less saline ones. Significant and accelerated changes in hyporegulation ability evolved in parallel with smaller and more progressive increases in desiccation resistance across the phylogeny, associated with the colonization of meso- and hypersaline waters during global aridification events. All species with high hyporegulation ability were also desiccation-resistant, but not vice versa. Overall, results are consistent with the hypothesis that desiccation resistance mechanisms evolved first and provided the physiological basis for the development of hyporegulation ability, allowing these insects to colonize and diversify across meso- and hypersaline habitats.

Pleistocene range shifts, refugia and the origin of widespread species in western Palaearctic water beetles.

Quaternary glacial cycles drove major shifts in both the extent and location of the geographical ranges of many organisms. During glacial maxima, large areas of central and northern Europe were inhospitable to temperate species, and these areas are generally assumed to have been recolonized during interglacials by range expansions from Mediterranean refugia. An alternative is that this recolonization was from non-Mediterranean refugia, in central Europe or western Asia, but data on the origin of widespread central and north European species remain fragmentary, especially for insects. We studied three widely distributed lineages of freshwater beetles (the Platambus maculatus complex, the Hydraena gracilis complex, and the genus Oreodytes), all restricted to running waters and including both narrowly distributed southern endemics and widespread European species, some with distributions spanning the Palearctic. Our main goal was to determine the role of the Pleistocene glaciations in shaping the diversification and current distribution of these lineages. We sequenced four mitochondrial and two nuclear genes in populations drawn from across the ranges of these taxa, and used Bayesian probabilities and Maximum Likelihood to reconstruct their phylogenetic relationships, age and geographical origin. Our results suggest that all extant species in these groups are of Pleistocene origin. In the H. gracilis complex, the widespread European H. gracilis has experienced a rapid, recent range expansion from northern Anatolia, to occupy almost the whole of Europe. However, in the other two groups widespread central and northern European taxa appear to originate from central Asia, rather than the Mediterranean. These widespread species of eastern origin typically have peripherally isolated forms in the southern Mediterranean peninsulas, which may be remnants of earlier expansion-diversification cycles or result from incipient isolation of populations during the most recent Holocene expansion. The accumulation of narrow endemics of such lineages in the Mediterranean may result from successive cycles of range expansion, with subsequent speciation (and local extinction in glaciated areas) through multiple Pleistocene climatic cycles.

Physiological niche and geographical range in European diving beetles (Coleoptera: Dytiscidae).

Geographical ranges vary greatly in size and position, even within recent clades, but the factors driving this remain poorly understood. In aquatic beetles, thermal niche has been shown to be related to both the relative range size and position of congeners but whether other physiological parameters play a role is unknown. Metabolic plasticity may be critical for species occupying more variable thermal environments and maintaining this plasticity may trade-off against other physiological processes such as immunocompetence. Here we combine data on thermal physiology with measures of metabolic plasticity and immunocompetence to explore these relationships in Deronectes (Dytiscidae). While variation in latitudinal range extent and position was explained in part by thermal physiology, aspects of metabolic plasticity and immunocompetence also appeared important. Northerly distributed, wide-ranging species apparently used different energy reserves under thermal stress from southern endemic congeners and differed in their antibacterial defences. This is the first indication that these processes may be related to geographical range, and suggests parameters that may be worthy of exploration in other taxa.

Oxygen-limited thermal tolerance is seen in a plastron-breathing insect and can be induced in a bimodal gas exchanger.

Thermal tolerance has been hypothesized to result from a mismatch between oxygen supply and demand. However, the generality of this hypothesis has been challenged by studies on various animal groups, including air-breathing adult insects. Recently, comparisons across taxa have suggested that differences in gas exchange mechanisms could reconcile the discrepancies found in previous studies. Here, we test this suggestion by comparing the behaviour of related insect taxa with different gas exchange mechanisms, with and without access to air. We demonstrate oxygen-limited thermal tolerance in air-breathing adults of the plastron-exchanging water bug Aphelocheirus aestivalis. Ilyocoris cimicoides, a related, bimodal gas exchanger, did not exhibit such oxygen-limited thermal tolerance and relied increasingly on aerial gas exchange with warming. Intriguingly, however, when denied access to air, oxygen-limited thermal tolerance could also be induced in this species. Patterns in oxygen-limited thermal tolerance were found to be consistent across life-history stages in these insects, with nymphs employing the same gas exchange mechanisms as adults. These results advance our understanding of oxygen limitation at high temperatures; differences in the degree of respiratory control appear to modulate the importance of oxygen in setting tolerance limits.

New species and new records of Mesoceration Janssens, 1967 from South Africa (Coleoptera, Hydraenidae).

Mesoceration Janssens, 1967 is the most speciose genus of Prosthetopinae, most of the 44 described species being restricted to South Africa, and almost all occupying the benthic zone of streams and rivers. Here seven species are described as new: Mesoceration caniplenum sp. nov., M. foggoi sp. nov., M. helmei sp. nov., M. hirsutum sp. nov., M. rugulosum sp. nov., M. sewefonteinense sp. nov. and M. sinclairi sp. nov., bringing the number of known species to 51. All seven new species have been discovered during recent, targeted sampling of South African Hydraenidae. New collection records resulting from this fieldwork are also provided for 27 previously described species, together with ecological notes.

Thermal niche evolution and geographical range expansion in a species complex of western Mediterranean diving beetles.

BACKGROUND: Species thermal requirements are one of the principal determinants of their ecology and biogeography, although our understanding of the interplay between these factors is limited by the paucity of integrative empirical studies. Here we use empirically collected thermal tolerance data in combination with molecular phylogenetics/phylogeography and ecological niche modelling to study the evolution of a clade of three western Mediterranean diving beetles, the Agabus brunneus complex. RESULTS: The preferred mitochondrial DNA topology recovered A. ramblae (North Africa, east Iberia and Balearic islands) as paraphyletic, with A. brunneus (widespread in the southwestern Mediterranean) and A. rufulus (Corsica and Sardinia) nested within it, with an estimated origin between 0.60-0.25 Ma. All three species were, however, recovered as monophyletic using nuclear DNA markers. A Bayesian skyline plot suggested demographic expansion in the clade at the onset of the last glacial cycle. The species thermal tolerances differ significantly, with A. brunneus able to tolerate lower temperatures than the other taxa. The climatic niche of the three species also differs, with A. ramblae occupying more arid and seasonal areas, with a higher minimum temperature in the coldest month. The estimated potential distribution for both A. brunneus and A. ramblae was most restricted in the last interglacial, becoming increasingly wider through the last glacial and the Holocene. CONCLUSIONS: The A. brunneus complex diversified in the late Pleistocene, most likely in south Iberia after colonization from Morocco. Insular forms did not differentiate substantially in morphology or ecology, but A. brunneus evolved a wider tolerance to cold, which appeared to have facilitated its geographic expansion. Both A. brunneus and A. ramblae expanded their ranges during the last glacial, although they have not occupied areas beyond their LGM potential distribution except for isolated populations of A. brunneus in France and England. On the islands and possibly Tunisia secondary contact between A. brunneus and A. ramblae or A. rufulus has resulted in introgression. Our work highlights the complex dynamics of speciation and range expansions within southern areas during the last glacial cycle, and points to the often neglected role of North Africa as a source of European biodiversity.

New species and new records of Pterosthetops: eumadicolous water beetles of the South African Cape (Coleoptera, Hydraenidae).

Pterosthetops is one of a number of hydraenid genera endemic to the Cape of South Africa, whose minute moss beetle fauna is amongst the most diverse on earth. Here seven species are described as new: Pterosthetops baini sp. nov., Pterosthetops coriaceus sp. nov., Pterosthetops indwei sp. nov., Ptersothetops pulcherrimus sp. nov., Pterosthetops swartbergensis sp. nov., Pterosthetops tuberculatus sp. nov. and Pterosthetops uitkyki sp. nov., all from mountains in the Western Cape region. New collection records are also provided for all five previously described members of the genus, together with a revised key. Pterosthetops appear to be specialist inhabitants of seepages over rock faces (hygropetric/madicolous habitats), rarely being found outside such situations.

A distinctive new species of Protozantaena Perkins, 1997 from lowland Namaqualand, South Africa (Coleoptera, Hydraenidae).

Protozantaena gigantea sp. nov. is described, based on specimens collected from residual pools in a drying seasonal river in Namaqualand, Northern Cape Province, South Africa. Morphologically, the new species appears to be related to P. labrata Perkins, 1997 from Namibia and P. birdi Bilton, 2022, from the Great Escarpment in the Eastern Cape Province. At up to 2.0 mm in body length, the new species, whilst small, is by far the largest African Protozantaena Perkins, 1997 known to date. The opportunity is also taken to report a new record for P. birdi in the Eastern Cape Drakensberg.

Nuanced impacts of the invasive aquatic plant Crassula helmsii on Northwest European freshwater macroinvertebrate assemblages.

Invasive alien species are considered one of the greatest threats to global biodiversity, and are particularly problematic in aquatic systems. Given the foundational role of macrophytes in most freshwaters, alien aquatic plant invasions may drive strong bottom-up impacts on recipient biota. Crassula helmsii (New Zealand pygmyweed) is an Australasian macrophyte, now widespread in northwest Europe. Crassula helmsii rapidly invades small lentic waterbodies, where it is generally considered a serious threat to native biodiversity. The precise ecological impacts of this invasion remain poorly understood, however, particularly with respect to macroinvertebrates, which comprise the bulk of freshwater faunal biodiversity. We conducted a field study of ponds, ditches and small lakes across the core of C. helmsii's invasive range (United Kingdom, Belgium and the Netherlands), finding that invaded sites had higher macroinvertebrate taxon richness than uninvaded sites, and that many infrequent and rare macroinvertebrates co-occurred with C. helmsii. Alien macroinvertebrates were more abundant in C. helmsii sites, however, particularly the North American amphipod Crangonyx pseudogracilis. At the order level, water beetle (Coleoptera) richness and abundance were higher in C. helmsii sites, whereas true fly (Diptera) abundance was higher in uninvaded sites. Taxonomic and functional assemblage composition were both impacted by invasion, largely in relation to taxa and traits associated with detritivory, suggesting that the impacts of C. helmsii on macroinvertebrates are partly mediated by the availability and palatability of its detritus. The nuanced effects of C. helmsii on macroinvertebrates found here should encourage further quantitative research on the impacts of this invasive plant, and perhaps prompt a more balanced re-evaluation of its effects on native aquatic macrofauna.

The effect of geographical scale of sampling on DNA barcoding.

Eight years after DNA barcoding was formally proposed on a large scale, CO1 sequences are rapidly accumulating from around the world. While studies to date have mostly targeted local or regional species assemblages, the recent launch of the global iBOL project (International Barcode of Life), highlights the need to understand the effects of geographical scale on Barcoding's goals. Sampling has been central in the debate on DNA Barcoding, but the effect of the geographical scale of sampling has not yet been thoroughly and explicitly tested with empirical data. Here, we present a CO1 data set of aquatic predaceous diving beetles of the tribe Agabini, sampled throughout Europe, and use it to investigate how the geographic scale of sampling affects 1) the estimated intraspecific variation of species, 2) the genetic distance to the most closely related heterospecific, 3) the ratio of intraspecific and interspecific variation, 4) the frequency of taxonomically recognized species found to be monophyletic, and 5) query identification performance based on 6 different species assignment methods. Intraspecific variation was significantly correlated with the geographical scale of sampling (R-square = 0.7), and more than half of the species with 10 or more sampled individuals (N = 29) showed higher intraspecific variation than 1% sequence divergence. In contrast, the distance to the closest heterospecific showed a significant decrease with increasing geographical scale of sampling. The average genetic distance dropped from > 7% for samples within 1 km, to < 3.5% for samples up to > 6000 km apart. Over a third of the species were not monophyletic, and the proportion increased through locally, nationally, regionally, and continentally restricted subsets of the data. The success of identifying queries decreased with increasing spatial scale of sampling; liberal methods declined from 100% to around 90%, whereas strict methods dropped to below 50% at continental scales. The proportion of query identifications considered uncertain (more than one species < 1% distance from query) escalated from zero at local, to 50% at continental scale. Finally, by resampling the most widely sampled species we show that even if samples are collected to maximize the geographical coverage, up to 70 individuals are required to sample 95% of intraspecific variation. The results show that the geographical scale of sampling has a critical impact on the global application of DNA barcoding. Scale-effects result from the relative importance of different processes determining the composition of regional species assemblages (dispersal and ecological assembly) and global clades (demography, speciation, and extinction). The incorporation of geographical information, where available, will be required to obtain identification rates at global scales equivalent to those in regional barcoding studies. Our result hence provides an impetus for both smarter barcoding tools and sprouting national barcoding initiatives-smaller geographical scales deliver higher accuracy.

Respiratory control in aquatic insects dictates their vulnerability to global warming.

Forecasting species responses to climatic warming requires knowledge of how temperature impacts may be exacerbated by other environmental stressors, hypoxia being a principal example in aquatic systems. Both stressors could interact directly as temperature affects both oxygen bioavailability and ectotherm oxygen demand. Insufficient oxygen has been shown to limit thermal tolerance in several aquatic ectotherms, although, the generality of this mechanism has been challenged for tracheated arthropods. Comparing species pairs spanning four different insect orders, we demonstrate that oxygen can indeed limit thermal tolerance in tracheates. Species that were poor at regulating oxygen uptake were consistently more vulnerable to the synergistic effects of warming and hypoxia, demonstrating the importance of respiratory control in setting thermal tolerance limits.

Hydraena lotti sp. nov., a new member of the "Haenydra" lineage from the Peloponnese (Greece), with additional records of Hydraena species in the region (Coleoptera, Hydraenidae).

Hydraena lotti sp. nov. (Coleoptera, Hydraenidae) is described from the southern Peloponnese, Greece; the 92nd known member of the "Haenydra" lineage. The new species belongs to the H. emarginata complex, being closest morphologically to Hydraena pelops Jäch, 1995, from the south and east of the Taygetos range, H. pangaei Jäch, 1992, endemic to Mount Pangaeon in northeastern Greece, and H. samnitica Fiori, 1904, from central Italy. Characters on which the species can be distinguished are discussed; male genitalia and female elytral apices being particularly diagnostic. The ecology of H. lotti is described in the context of other members of the genus in the region. To date, the new species has only been found in small headwater streams at altitudes above 1,000 m in the northwest of the Taygetos range, where it can, however, be locally frequent. The opportunity is taken to provide an updated checklist of Peloponnese "Haenydra", together with new distributional records of selected Hydraena species, including H. arachthi Ferro & Jäch, 2000, which is reported from the peninsula for the first time.

Description of the male of Sebasthetops omaliniformis Jäch, 1998--a phylogenetically isolated water beetle from South Africa, with notes on its ecology (Coleoptera, Hydraenidae).

Sebasthetops omaliniformis Jäch, 1998, is a morphologically aberrant hydraenid, known from two female specimens collected from the Western Cape of South Africa in 1988. Recent fieldwork has resulted in the rediscovery of the species, close to the type locality. The male of S. omaliniformis is described from this material, and the opportunity taken to publish a record of Sebasthetops from the Langeberg, where it was collected in 1979. S. omaliniformis lives in deep water riffles in the upper reaches of mountain streams, and is strongly brachypterous. Like a number of other running-water insects known from the Cape fold mountains the species appears to have a narrow geographical and ecological range, deserving of high conservation status.

Prosthetops wolybergensis sp. nov.--a giant amongst the 'minute moss beetles', with new data on other members of the genus (Coleoptera, Hydraenidae).

Prosthetops wolfbergensis sp. nov. (Coleoptera, Hydraenidae) is described from the Western and Northern Cape regions of South Africa; the new species, at up to 4.2 mm total length, apparently being the longest 'minute moss beetle' described to date. Prosthetops species are characteristic inhabitants of temporary rainwater pools and seepages on exposed plateau in the Cape fold mountains, and on rock outcrops beside streams and rivers. New collection records and ecological data are given for members of the genus, and the female of Prosthetops pronotus Perkins & Balfour-Browne, 1994 is described and illustrated for the first time.

A taxonomic revision of South African Sharphydrus, with the description of two new species (Coleoptera: Dytiscidae: Bidessini).

Sharphydrus Omer-Cooper, 1958 is one of two endemic bidessine genera currently recognised from South Africa. Here Sharphydrus brincki sp. nov. and Sharphydrus kamiesbergensis sp. nov. are described from the Cederberg and Gydopas areas of the Western Cape, and the high Kamiesberg of the Northern Cape respectively, doubling the known species of this genus. It is shown that S. brincki sp. nov. has been included under S. capensis (Omer-Cooper, 1955) in the past, but that these are quite distinct taxa, differing in the extent of their elytral keels and male genitalia. Sharphydrus species are inhabitants of pools in seasonally fluctuating rivers, the new species described here occurring in areas which are somewhat transitional between fynbos and karoo biomes. An updated key is presented to Sharphydrus species, together with data on the distribution and ecology of known species, and a discussion of the status of the genus within the Bidessini.