Philippa Kaur

Department of Zoology, University of Cambridge, Cambridge, UKYucheng Wang, Bianca De Sanctis, Ruairidh Macleod, Daniel Money & Eske WillerslevLundbeck Foundation GeoGenetics Centre, Globe Institute, University of Copenhagen, Copenhagen, DenmarkYucheng Wang, Ana Prohaska, Jialu Cao, Antonio Fernandez-Guerra, James Haile, Kurt H. Kjær, Thorfinn Sand Korneliussen, Nicolaj Krog Larsen, Ruairidh Macleod, Hugh McColl, Mikkel Winther Pedersen, Fernando Racimo, Alexandra Rouillard, Anthony H. Ruter, Lasse Vinner, David J. Meltzer & Eske WillerslevALPHA, State Key Laboratory of Tibetan Plateau Earth System, Environment and Resources (TPESER), Institute of Tibetan Plateau Research (ITPCAS), Chinese Academy of Sciences (CAS), Beijing, ChinaYucheng WangKey Laboratory of Western China’s Environmental Systems (Ministry of Education), College of Earth and Environmental Science, Lanzhou University, Lanzhou, ChinaHaoran DongGénomique Métabolique, Genoscope, Institut François Jacob, CEA, CNRS, Univ Evry, Université Paris-Saclay, Evry, FranceAdriana Alberti, France Denoeud & Patrick WinckerInstitute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, CEA, CNRS, Gif-sur-Yvette, FranceAdriana AlbertiThe Arctic University Museum of Norway, UiT—The Arctic University of Norway, Tromsø, NorwayInger Greve Alsos, Eric Coissac, Galina Gusarova, Youri Lammers & Marie Kristine Føreid MerkelDepartment of Geography and Environment, University of Hawaii, Honolulu, HI, USADavid W. BeilmanDepartment of Geosciences and Natural Resource Management, University of Copenhagen, Copenhagen, DenmarkAnders A. BjørkInstitute of Earth Sciences, St Petersburg State University, St Petersburg, RussiaAnna A. Cherezova & Grigory B. FedorovArctic and Antarctic Research Institute, St Petersburg, RussiaAnna A. Cherezova & Grigory B. FedorovUniversité Grenoble-Alpes, Université Savoie Mont Blanc, CNRS, LECA, Grenoble, FranceEric CoissacDepartment of Genetics, University of Cambridge, Cambridge, UKBianca De Sanctis & Richard DurbinCarlsberg Research Laboratory, Copenhagen V, DenmarkChristoph Dockter & Birgitte SkadhaugeSchool of Geography and Environmental Science, University of Southampton, Southampton, UKMary E. EdwardsAlaska Quaternary Center, University of Alaska Fairbanks, Fairbanks, AK, USAMary E. EdwardsSchool of Environment, Earth and Ecosystem Sciences, The Open University, Milton Keynes, UKNeil R. Edwards & Philip B. HoldenCenter for the Environmental Management of Military Lands, Colorado State University, Fort Collins, CO, USAJulie EsdaleDepartment of Earth and Atmospheric Sciences, University of Alberta, Edmonton, Alberta, CanadaDuane G. FroeseFaculty of Biology, St Petersburg State University, St Petersburg, RussiaGalina GusarovaDepartment of Glaciology and Climate, Geological Survey of Denmark and Greenland, Copenhagen K, DenmarkKristian K. KjeldsenDepartment of Earth Science, University of Bergen, Bergen, NorwayJan Mangerud & John Inge SvendsenBjerknes Centre for Climate Research, Bergen, NorwayJan Mangerud & John Inge SvendsenDepartment of Geology, Quaternary Sciences, Lund University, Lund, SwedenPer MöllerCenter for Macroecology, Evolution and Climate, Globe Institute, University of Copenhagen, Copenhagen Ø, DenmarkDavid Nogués-Bravo, Hannah Lois Owens & Carsten RahbekCentre d’Anthropobiologie et de Génomique de Toulouse, Faculté de Médecine Purpane, Université Paul Sabatier, Toulouse, FranceLudovic OrlandoCenter for Global Mountain Biodiversity, Globe Institute, University of Copenhagen, Copenhagen, DenmarkHannah Lois Owens & Carsten RahbekGates of the Arctic National Park and Preserve, US National Park Service, Fairbanks, AK, USAJeffrey T. RasicDepartment of Geosciences, UiT—The Arctic University of Norway, Tromsø, NorwayAlexandra RouillardZoological Institute, Russian academy of sciences, St Petersburg, RussiaAlexei TikhonovResource and Environmental Research Center, Chinese Academy of Fishery Sciences, Beijing, ChinaYingchun XingCollege of Plant Science, Jilin University, Changchun, Jilin, ChinaYubin ZhangDepartment of Anthropology, Southern Methodist University, Dallas, TX, USADavid J. MeltzerWellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge, UKEske WillerslevMARUM, University of Bremen, Bremen, GermanyEske WillerslevAll authors contributed to the conception of the presented ideas. Y.W. and H.D. analysed the data. Y.W., D.J.M., A.P. and E.W. wrote the paper with inputs from all authors. More

Suborder Glossiphoniiformes Tessler & de Carle, 2018Family Glossiphoniidae Vaillant, 1890Comments. Our two-locus phylogeny reveals the presence of two large clades, corresponding to the subfamilies Glossiphoniinae and Haementeriinae (Fig. 1). The subfamily Theromyzinae Sawyer, 1986, delineated by some authors2,10,22, was not supported as a distant phylogenetic clade and their representatives are clustered within the monophyletic Glossiphoniinae. The same pattern was recovered by earlier phylogenetic reconstructions3,30,33,34. These data indicate that Theromyzinae may represent a synonym of the latter subfamily. However, a subfamily-level revision of the Glossiphoniidae is beyond the framework of the present study.Subfamily Glossiphoniinae Vaillant, 1890Genus Alboglossiphonia Lukin, 1976Type species: Alboglossiphonia heteroclita (Linnaeus, 1761) (= Hirudo heteroclita Linnaeus, 1761; by original designation).Arctic occurrences. Our results reveal that members of this genus are not common inhabitants of the Arctic but two species, A. heteroclita (Linnaeus, 1761) and A. sibirica sp. nov., cross the Arctic Circle on the Yamal Peninsula through the Ob and Taz rivers (Table 1). Previously, it was shown that A. heteroclita occurs in the lower Ob Basin, northern edge of Western Siberia23.Comments. This genus contains inconspicuous minute leeches and is characterized by a nearly global distribution1. It definitely requires an integrative taxonomic revision. Available genetic evidence (Fig. 1 and Supplementary Fig. S1) reveals that the North American populations of what was traditionally assigned to A. heteroclita should be considered a separate species, A. pallida (Verrill, 1872) (type locality: West River near New Haven, Connecticut, USA)35,36. Other species, which occurs in Siberia and the Far East, was tentatively assigned to Alboglossiphonia cf. papillosa (Braun, 1805) based on a darker pigmentation of its dorsum37,38 but it represents a separate North Asian species, which is described here.
Alboglossiphonia sibirica Bolotov, Eliseeva, Klass & Kondakov sp. nov = Alboglossiphonia heteroclita Lukin (1957): 27339 (identification error). = Alboglossiphonia heteroclita papillosa Kaygorodova et al. (2014): 337; Kaygorodova (2015): 4140 (identification error). = Alboglossiphonia cf. papillosa Klass et al. (2018): 2638 (identification error).Figures 4a, 5a, 7a, Supplementary Figs. S2a, S3a, S4, Supplementary Table S2.LSID: https://zoobank.org/urn:lsid:zoobank.org:act:19B581C3-E912-487C-B9EC-8E50DDEFD380.Holotype. RMBH Hir_0542_2-H (non-sequenced), RUSSIA: Lake Torfyanka, 43.0761° N, 131.9620° E, Vladivostok, Primorye, August 12, 2021, Y. E. Chapurina leg.Paratypes (N = 13). RUSSIA: 1 specimen RMBH Hir_0542_2 (sequenced: COI sequence acc. No. ON873332), the type locality, the same date, and collector; 1 specimen RMBH Hir_0396 (non-sequenced), an oxbow lake of Taz River, near Tazovsky settlement, 67.5063° N, 78.6751° E, Yamal-Nenets Region, August 22, 2019, E. S. Babushkin leg.; 1 specimen RMBH Hir_0394 (DNA voucher; sequenced: COI sequence acc. No. ON548508), Vitim River, 57.2010° N, 116.4300° E, Lena River basin, Vitimsky Nature Reserve, Irkutsk Region, July 12, 2019, E. S. Babushkin leg.; 4 specimens RMBH Hir_0013 (3 sequenced with DNA vouchers and one placed on 36 permanent slides as a series of slices; COI sequence acc. No. MH286267, MH286268, and MH286269; 18S rRNA sequence acc. No. MH286273), between zooids of a bryozoan colony, small floodplain lake of the Lena River near Yakutsk, 62.3076° N, 129.8999° E, Yakutia Republic, August 20, 2017, I. N. Bolotov leg.; 1 specimen RMBH Hir_0417_2 (DNA voucher; sequenced: COI sequence acc. No. ON548511), Oron Lake, Gnilaya Kurya Bay, 57.1750° N, 116.4031° E, Lena River basin, Vitimsky Nature Reserve, Irkutsk Region, July 1, 2019, E. S. Babushkin leg.; 1 specimen RMBH Hir_0409_1 (sequenced: COI sequence acc. No. ON548509), a roadside ditch in Knevichi settlement, 43.3886° N, 132.1880° E, Primorye, September 10, 2020, O. V. Aksenova et al. leg.; 1 specimen RMBH Hir_0413 (sequenced: COI sequence acc. No. ON548510), a puddle near railway at Artem city, 43.3794° N, 132.2188° E, Primorye, September 10, 2020, O. V. Aksenova et al. leg.; 1 specimen RMBH Hir_0003_3 (DNA voucher; sequenced: COI sequence acc. No. MN393256), Tumnin River, 49.9451° N, 139.9181° E, Khabarovsk Region, July 14, 2014, I. N. Bolotov & I. V. Vikhrev leg.; 1 specimen RMBH Hir_0509_1 (sequenced: COI sequence acc. No. ON548516), a reservoir on the Bolshoy Alim River, near Tolstovka settlement, 50.1981° N, 127.9431° E, Amur Region, July 3, 2021, O. V. Aksenova et al. leg.; 1 specimen RMBH Hir_0510_1 (DNA voucher; sequenced: COI sequence acc. No. ON548517), an oxbow lake of Bureya River, near Novospassk, 49.6756° N, 129.7343° E, Amur Region, July 3, 2021, O. V. Aksenova et al. leg.Etymology. The name of this species reflects its broad distribution in Siberia.Differential diagnosis. Small leech, which could be distinguished from other congeners by a combination of the following characters: dorsum covered by numerous small, shallow, and indistinct papillae, light yellow, with multiple dark spots and short dashes arranged to 18–24 longitudinal rows; these spots and dashes merged into longitudinal lines in the anterior half of the body (the dark markings pattern often lost in ethanol-preserved animals). Externally, the new species is similar to A. heteroclita, A. hyalina (O. F. Müller, 1773), and A. striata (Apáthy, 1888). However, all these species do not have numerous dark spots and short dashes arranged to multiple longitudinal rows. Additionally, A. heteroclita differs from the new species by having a median row of segmentally arranged dark spots and a smooth dorsum without papillae. A. hyalina differs from A. sibirica sp. nov. by the general lack of dark pigmentation. A. striata differs from the new species by having a median row of segmentally arranged dark transverse stripes and a smooth dorsum without papillae.Molecular diagnosis. The new species represents a separate genetic lineage but is more closely related to A. heteroclita (mean pairwise COI p-distance = 5.1%; range 4.9–5.4%). The intraspecific pairwise COI p-distance ranges from 0.0 to 2.1% (mean ± s.e.m. = 1.31 ± 0.10%; N = 14 sequences and 91 pairwise distance values). The GenBank acc. numbers of reference DNA sequences (COI and 18S rRNA) are given in Supplementary Table S2 and Supplementary Datasets S1–S2.Description. Small leech (body length up to 11.9 mm). Measurements of the type series are given in Supplementary Table S2. Body broad, leaf-like, ovate. Dorsum with numerous small, shallow, and indistinct papillae. Posterior sucker small, circular (maximum diameter of 2.25 mm), ventrally directed. Proboscis pore in the center of anterior sucker. Coloration of living animals: body dirty yellow with multiple brown spots and dashes arranged to longitudinal rows; in the anterior half of the body, these spots and dashes merged into longitudinal lines. Coloration of ethanol-preserved animals: body light yellow; dorsum with multiple dark spots and short dashes arranged to 18–24 longitudinal rows; these spots and dashes merged into longitudinal lines in the anterior half of the body but the dark markings pattern often lost due to preservation. Three pairs of eyespots; the eyespots of the distal pair joined into a single spot; the eyespots of the next two pairs are spaced apart and fused together. Venter light yellow or whitish. Total number of annuli: 70. Somites I–IV joined to form a head region, somites V–XXIV triannulate, somites XXV–XXVII uniannulate. Gonopores joined and open in the furrow XIIa1/a2. Reproductive system: 6 pairs of large, bag-like testisacs inter-segmentally from XIII/XIV to XIX/XX; atrium small, spherical, the atrial cornua twisted anteriorly; paired ejaculatory ducts twisted, short; paired ovisacs narrow, very short. Digestive system: proboscis sheath massive, long, thick; salivary glands diffuse; crop with 6 pairs of crop caeca: 1st-5th uniform, bag-like, 6th pair (posterior caeca) with 3 blind processes; intestine with 4 pairs of rather short processes and an ovate extention after the last pair of processes.Distribution. North Asia: Western Siberia, Eastern Siberia, the Russian Far East, and Mongolia39.Habitats and ecology. This species is known to occur in a broad range of freshwater environments such as rivers, oxbow lakes, large to small natural lakes, reservoirs, road ditches, and even puddles (Supplementary Dataset S2). An unusual example of its association with a bryozoan species was described from Eastern Siberia38. Probably, the record of an Alboglossiphonia leech in the mantle cavity of an unidentified lymnaeid snail from the Altai Mountains, Russia41 could also be attributed to this species. The life cycle of the new species is unknown.Genus Glossiphonia Johnson, 1816Type species: Glossiphonia complanata (Linnaeus, 1758) (= Hirudo complanata Linnaeus, 1758; by subsequent designation).Arctic occurrences. Representatives of this genus are the most remarkable component of the Arctic Glossiphoniidae fauna. Altogether seven species were recorded north of the Arctic Circle, two of which are new to science and described here (Table 1).Comments. In general, sequenced representatives of the genus Glossiphonia could phylogenetically be delineated to three species groups (or subgenera): (1) the complanata-group (= subgenus Glossiphonia s. str.); (2) the verrucata-group (= subgenus Boreobdella Johansson, 1929; type species: Clepsine verrucata Müller, 1844); and (3) the concolor-group (= subgenus Paratorix Lukin & Epstein, 1960; type species: Torix baicalensis Stschegolew, 1922) (Table 1, Fig. 1 and Supplementary Fig. S1).
Glossiphonia arctica Bolotov, Eliseeva, Klass & Kondakov sp. novFigures 4B, 5b,c, 7c, Supplementary Figs. S2b, S3b, S5, Supplementary Table S2.LSID: https://zoobank.org/urn:lsid:zoobank.org:act:FADF0993-A946-413A-9680-25BA0F9BE90D.Holotype. RMBH Hir_0457_2_1-H (sequenced: COI sequence acc. No. ON810735; 18S rRNA sequence acc. No. ON819028), RUSSIA: a large lake near Sob’ railway station, 67.0480°N, 65.6316°E, Polar Urals, June 23, 2021, A. V. Kondakov et al. leg.Paratypes (N = 18). 18 specimens RMBH Hir_0457 (two specimens sequenced: COI sequence acc. No. ON810736 and ON810737; 18S rRNA sequence acc. No. ON819029; one specimen placed on 18 permanent slides as a series of slices), the type locality, the same date, and collectors.Etymology. The name of the new species indicates that its type locality is situated in the Arctic Region.Differential diagnosis. Medium-sized leech, which could be distinguished from other congeners by a combination of the following characters: dorsum with four rows of ovate, broad but very shallow and indistinct papillae on annulus a2 (outer paramedian and inner paramarginal series); each papilla bears ovate light yellow or white spot; dorsal black markings pattern absent. Externally, the new species is similar to G. mollissima. However, the latter species differs from G. arctica sp. nov. by having larger papillae and a well-developed black markings pattern dorsally.Molecular diagnosis. The new species represents a separate genetic lineage belonging to the verrucata-group (Fig. 1). The pairwise COI p-distance of the new species from other congeners varies from 7.0 to 12.4%. The intraspecific pairwise COI p-distance ranges from 0.0 to 0.2% (mean ± s.e.m. = 0.10 ± 0.05%; N = 3 sequences and 3 pairwise distance values). The GenBank acc. numbers of reference DNA sequences (COI and 18S rRNA) are given in Supplementary Table S2 and Supplementary Datasets S1–S2.Description. Medium-sized leech (body length up to 13.3 mm). Measurements of the type series are given in Supplementary Table S2. Body broad, leaf-like, ovate. Dorsum with four rows of ovate, broad but very shallow and indistinct papillae on annulus a2 (outer paramedian and inner paramarginal series). Posterior sucker small, circular (maximum diameter of 1.9 mm), ventrally directed. Proboscis pore in the center of anterior sucker. Coloration of living animals: body almost transparent, light brown, with multiple yellowish pigment cells. Coloration of ethanol-preserved animals: dorsum beige to light brown, with darker broad inner paramedian lines and light yellowish areas laterally and anteriorly; ovate light yellow or white spots at each papillae on annulus a2 arranged into four longitudinal rows (outer paramedian and inner paramarginal), sometimes with a few white spots between them. Three pairs of ovate eyespots arranged to two parallel rows; in some specimens eyes on each side are joined to a single large spot. Venter whitish to light brown, sometimes with irregular brownish shading. Total number of annuli: 70. Somites I–III uniannulate, IV biannulate, V–XXIV triannulate, XXV biannulate, XXVII uniannulate. The male and female genital pores are separated by two annuli and are located in the furrows XIa3/XIIa1 and XIIa2/a3, respectively. Reproductive system: 6 pairs of spherical testisacs inter-segmentally from XIII/XIV to XVIII/XIX; atrium spherical, the atrial cornua large, twisted anteriorly; paired ejaculatory ducts very long, extending to XVIII; paired ovisacs massive, long, with multiple lobes, arranged as loops, extending to XVIII (pregnant specimen with eggs). Digestive system: proboscis sheath massive, thick, elongated; esophagus narrow; salivary glands diffuse; crop with 7 pairs of crop caeca: 1st-6th uniform, bag-like, 7th pair (posterior caeca) with 4 blind processes and several smaller lobes; intestine enlarged, with 4 pairs of large, long, bag-like processes, expanding distally, each with several short lobes; a large circular extension after the last pair of processes.Distribution. Polar Urals (not known beyond the type locality).Habitats and ecology. The type series of this species was collected from a natural mountain lake with stony bottom. The leeches were recorded beneath flat stones (Fig. 3b); their feeding behavior and life cycle remain unknown.
Glossiphonia taymyrensis Bolotov, Eliseeva, Klass & Kondakov sp. novFigures 4E, 5d, 7b, Supplementary Figs. S2h, S3c, S6, Supplementary Table S2.LSID: https://zoobank.org/urn:lsid:zoobank.org:act:40269BF4-FE1C-4269-A7CC-41020789DC44.Holotype. RMBH Hir_0258_1-H (sequenced: COI sequence acc. No. ON810695), RUSSIA: small lake near Dudinka on Taymyr Peninsula, 69.4008°N, 86.3384°E, July 16, 2018, O. V. Aksenova et al. leg.Paratypes (N = 8). RUSSIA: 2 specimens RMBH Hir_0263_1 and RMBH Hir_0264_3 (sequenced: COI sequence acc. No. ON810701 and ON810705; 18S rRNA sequence acc. No. ON819017), the type locality, the same date, and collectors; 2 specimens RMBH Hir_0256_1 (one sequenced and one placed on 20 permanent slides as a series of slices; COI sequence acc. No. ON810693), small lake near Dudinka on Taymyr Peninsula, 69.3987° N, 86.3505° E, July 16, 2018, O. V. Aksenova et al. leg.; 1 specimen RMBH Hir_0261_2 (sequenced: COI sequence acc. No. ON810699; 18S rRNA sequence acc. No. ON819016), small lake near Dudinka on Taymyr Peninsula, 69.4014° N, 86.3250° E, July 16, 2018, O. V. Aksenova et al. leg.; 1 specimen RMBH Hir_0265_2 (sequenced: COI sequence acc. No. ON810706; 18S rRNA sequence acc. No. ON819021), Bolgokhtokh River near Dudinka, Taymyr Peninsula, 69.3780° N, 87.2215° E, July 21, 2018, O. V. Aksenova et al. leg.; 1 specimen RMBH Hir_0488 (sequenced: COI sequence acc. No. ON810755), a lake on Putorana Plateau, 68.7607° N, 91.9014° E, July, 2021, E. S. Chertoprud leg.; 1 specimen RMBH Hir_0449 (sequenced: COI sequence acc. No. ON810731), Pyzas River near Ust-Kabyrza settlement, 52.8277° N, 88.3973° E, Tashtagolsky District, Kemerovo Region, July 23, 2020, E. S. Babushkin & M. V. Vinarski leg.Etymology. The new species is named after the Taymyr Peninsula, where the majority of the type specimens were collected.Differential diagnosis. Small leech with broad, leaf-like, ovate body; three pairs of eyespots (distal pair joined; next two pairs separate); dorsal papillae absent; dorsal coloration with two inner paramedian rows of black spots, sometimes joining into unclear dashed lines; two annuli between the male (XIa3/XIIa1) and female (XIIa2/a3) genital pores. The new species largely resembles G. complanata but could be distinguished from it by having a smooth dorsum, without clear papillae. These taxa seem to have non-overlapping, allopatric ranges and, hence, could be separated on the basis of geographic criteria. However, the DNA approach seems to be the most appropriate way to distinguish these two species.Molecular diagnosis. The new species represents a separate genetic lineage belonging to the complanata-group (Fig. 1). The pairwise COI p-distance of the new species from other congeners varies from 6.0 to 12.2%. The intraspecific pairwise COI p-distance ranges from 0.0 to 1.1% (mean ± s.e.m. = 0.52 ± 0.07%; N = 8 sequences and 28 pairwise distance values). The GenBank acc. numbers of reference DNA sequences (COI and 18S rRNA) are given in Supplementary Table S2 and Supplementary Datasets S1–S2.Description. Small leech (body length up to 11.3 mm). Measurements of the type series are given in Supplementary Table S2. Body broad, leaf-like, ovate. Dorsum smooth, without clear papillae. Posterior sucker ovate (maximum diameter of 3.0 mm), ventrally directed. Proboscis pore in the center of anterior sucker. Coloration of living animals: not examined. Coloration of ethanol-preserved animals: (1) typical form having beige to light brown ground color without light spots but with darker brown coloration between inner paramedian lines; (2) melanic forms having dark brown ground color with four rows of large yellow spots (outer paramedian and marginal series) and yellow median stripe anteriorly (f. ‘maculosa’) or with strongly reduced yellow markings pattern. In all forms, there are two inner paramedian rows of black spots, sometimes joining into unclear dashed lines. Three pairs of ovate eyespots; the eyespots of the distal pair joined into a single spot; the eyespots of the next two pairs separate and are spaced apart. In the typical form, venter light yellow, with paired brown median and outer paramedian lines, which may be reduced to series of narrow brown longitudinal stripes. In melanic forms, ventral markings is more developed, with a series of brown longitudinal lines from median to inner paramarginal position and outer paramarginal brown spots. Posterior sucker with dense brown spots in melanic forms and with scarce brown spots in typical form. Total number of annuli: 68. Somites I–IV uniannulate, V–XXIV triannulate, XXV biannulate, XXVI–XXVII uniannulate. The male and female genital pores are separated by two annuli and are located in the furrows XIa3/XIIa1 and XIIa2/a3, respectively. Reproductive system: 6 pairs of spherical testisacs inter-segmentally from XII/XIII to XVIII/XIX; atrium ovate, the atrial cornua directed laterally; paired ejaculatory ducts twisted, short; paired ovisacs short, thick (undeveloped). Digestive system: salivary glands diffuse; proboscis sheath moderately thick; esophagus ovate; crop with 6 pairs of massive, bag-like, uniform crop caeca; intestine with 4 pairs of processes.Distribution. Western and Eastern Siberia.Habitats and ecology. The new species was recorded from natural lakes and rivers (Supplementary Dataset S2); its feeding behavior and life cycle are unknown.Genus Hyperboreomyzon Bolotov, Eliseeva, Klass & Kondakov gen. novLSID: https://zoobank.org/urn:lsid:zoobank.org:act:298FF41E-AF0D-4442-9F82-3022B8094A67.Type species: Hyperboreomyzon polaris gen. & sp. nov.Etymology. This name is compiled using two Greek words: ‘Hyperborea’ (meaning a mythical far northern land) and ‘myzon’ (meaning sucking).Diagnosis. Medium-sized, elongate, sub-fusiform glossiphoniid leeches; body and posterior sucker densely covered by shallow, ‘fish-scale’-like papillae; somite V biannulate; somites XII–XXIII secondarily sexannulate dorsally and ventrally due to the presence of very deep, prominent furrows separating each annulus to two semi-annuli; six rows of prominent dorsal tubercle-like papillae at a2 (inner paramedian, inner paramarginal, and marginal series) from V to XXVI; two pairs of circular eyespots on II and Va1 at inner paramedian position; gonopores at the furrows XIa3/XIIa1 (male) and XIIa2/a3 (female) and separated by two annuli; male atrium spherical; proboscis pore opens in a thick velar fold in the anterior half of oral sucker; one pair of compact, massive, elongated, incurved salivary glands, each gland with a bunch of a few short processes apically; 9 crop caeca pairs. Comparison of the new genus with other genera in the family based on morphological and anatomical features is presented in
Supplementary Table S3. Sexannulate condition was also recorded in the genus Actinobdella Moore, 1901 from North America36, but it differs from Hyperboreomyzon gen. nov. by having one pair of eyespots, diffuse salivary glands, and an apical position of proboscis pore (Supplementary Table S3).Comments. This genus is established for a single species, which is described below.
Hyperboreomyzon polaris Bolotov, Eliseeva, Klass & Kondakov gen. & sp. nov.Figures 4J, 5e, 6a-j, 7c, Supplementary Figs. S21, S8, S9, S10, S11, Supplementary Table S2.LSID: https://zoobank.org/urn:lsid:zoobank.org:act:503A9A26-CEDE-4747-952D-8416AE4EF4EB.Holotype. RMBH Hir_0486-H (sequenced: COI sequence acc. No. ON810753; 18S rRNA sequence acc. No. ON819030), RUSSIA: small alpine lake on Putorana Plateau, 68.9008°N, 94.1599°E, July, 2021, E. S. Chertoprud leg.Paratypes (N = 2). RUSSIA: 1 specimen RMBH Hir_0689 (dissected and placed on 60 permanent slides as a series of slices), small alpine lake on Putorana Plateau, 68.6659° N, 93.1365° E, August 11, 2021, E. S. Chertoprud leg.; 1 specimen RMBH Hir_0216 (sequenced and dissected; COI sequence acc. No. ON810677; 18S rRNA sequence acc. No. ON819005), water puddle on Kolguev Island, 68.9300° N, 49.0303° E, August 12, 2018, O. V. Travina & V. M. Spitsyn leg.Etymology. The name of the new species reflects its occurrences in polar (Arctic) areas of Eurasia.Differential diagnosis. As for the genus.Molecular diagnosis. None of congeneric species is known. Based on uncorrected pairwise COI p-distances between a haplotype of the new taxon and selected species-level haplotypes in each genus (Supplementary Table S1), Hyperboreomyzon seems to be more closely related to members of Hemiclepsis (mean distance ± s.e.m. = 11.62 ± 0.15%, range = 9.75–14.08%, N = 9) and Theromyzon (mean distance ± s.e.m. = 11.37 ± 0.07%, range = 10.47–12.64%, N = 9) without significant differences between distances from these two genera (Mann–Whitney test: p = 0.72). Other Glossiphoniidae genera are more distantly related, with a mean pairwise uncorrected COI p-distance of  > 13.0% (Mann–Whitney test: p  More

Author notesMikayla M. ClarkPresent address: University of Tennessee, Knoxville, TN, USAMichael W. SneddonPresent address: Predicine, Inc., Hayward, CA, USARoman SutorminPresent address: Google, Inc., San Francisco, CA, USAAuthors and AffiliationsLawrence Berkeley National Laboratory, Berkeley, CA, USADylan Chivian, Sean P. Jungbluth, Paramvir S. Dehal, Elisha M. Wood-Charlson, Richard S. Canon, Gavin A. Price, William J. Riehl, Michael W. Sneddon, Roman Sutormin & Adam P. ArkinOak Ridge National Laboratory, Oak Ridge, TN, USABenjamin H. Allen, Mikayla M. Clark, Miriam L. Land & Robert W. CottinghamArgonne National Laboratory, Lemont, IL, USATianhao Gu, Qizhi Zhang & Chris S. HenryAuthorsDylan ChivianSean P. JungbluthParamvir S. DehalElisha M. Wood-CharlsonRichard S. CanonBenjamin H. AllenMikayla M. ClarkTianhao GuMiriam L. LandGavin A. PriceWilliam J. RiehlMichael W. SneddonRoman SutorminQizhi ZhangRobert W. CottinghamChris S. HenryAdam P. ArkinCorresponding authorsCorrespondence to
Dylan Chivian or Adam P. Arkin. More

Rearing O. biroi pupae in social isolation and collecting pupal fluidIn O. biroi colonies, larvae and pupae develop in discrete and synchronized cohorts26. Ten days after the first larvae had entered pupation in a large stock colony, the entire colony was anaesthetized using a CO2 pad, and white pupae were separated using a paintbrush. Pupae were individually placed in 0.2 ml PCR tubes with open lid. These tubes were then placed inside 1.5 ml Eppendorf tubes with 5 µl sterile water at the bottom to provide 100% relative humidity. The outer tubes were closed and kept in a climate room at 25 °C. The inner tube in this design prevents the pupa from drowning in the water reservoir. The outer tubes were kept closed throughout the experiment, except for once a day when the tubes were opened to remove pupal social fluid. Pulled glass capillaries were prepared as described elsewhere29, and used to remove and/or collect secretion droplets. We were careful to leave no remains of the secretion behind on the pupae or the inside of the tubes. To ensure that all secretion had been removed, pupae were taken out of the tube after fluid collection and briefly placed on a tissue paper to absorb any excess liquid. The inner tubes were replaced if needed—for example, if fluid traces were visible on the old tube after collection. Each pupa was checked daily for secretion (absent or present), onset of melanization and eclosion, and whether the pupa was alive (responding to touch). Control groups of 30 pupae and 30 adult ants from the same stock colony and cohort as the isolated pupae were placed in Petri dishes with a plaster of Paris floor, and the same parameters as for the isolated pupae were scored daily. Experiments ended when all pupae had either eclosed or died. Newly eclosed (callow) workers moved freely inside the tube and showed no abnormalities when put in a colony. A pupa was declared dead if it did not shed its pupal skin and did not respond to touch three days after all pupae in the control group had eclosed.To calculate the average secretion volume per secreting pupa (Fig. 1d), the total volume collected daily from a group of isolated pupae (142–166 pupae) was divided by the number of pupae from which fluid had been collected that day. The total volume was determined by multiplying the height of the fluid’s meniscus in the capillary by πr², where r is the inner radius of the capillary (0.29 mm). While pupae were secreting, pupal whole-body wash samples were collected daily. The pupae were removed from colonies with adults and washed promptly with 1500 µl LC–MS grade water. Whole-body wash samples were lyophilized and reconstituted in 15 µl LC–MS grade water.Collecting additional ant species and honeybees, rearing pupae in social isolation, and collecting pupal fluidsColonies of the ants N. flavipes, T. sessile, P. pennsylvanica and Lasius neoniger were collected in NY state, USA (Central Park, Manhattan; Pelham Bay Park, Bronx; Prospect Park, Brooklyn; and Woodstock). Solenopsis invicta colonies were collected in Athens, GA, USA. M. mexicanus colonies were collected in Piñon Hills, CA, USA. Colonies comprised of queens, workers and brood were maintained in the laboratory in airtight acrylic boxes with plaster of Paris floors. Colonies were fed a diet of insects (flies, crickets and mealworms). White pupae were socially isolated, cocoons were removed in the case of P. pennsylvanica, and secretion droplets were collected from melanized pupae as described for O. biroi. A. mellifera pupae of unknown age were socially isolated from hive fragments (A&Z Apiaries, USA) and reared as described for O biroi, except that the rearing temperature was set to 32 °C. Relative humidity was set to either 100% to replicate conditions used for the different ant species, or to 75% as recommended in the literature30.Injecting dye and tracking pupal fluidInjection needles were prepared as in previous studies31. Injections were performed using an Eppendorf Femtojet with a Narishige micromanipulator. The Femtojet was set to Pi 1000 hPa and Pc 60 hPa. Needles were broken by gently touching the capillary tip to the side of a glass slide. To inject, melanized pupae were placed on ‘Sticky note’ tape (Post-it), with the abdomen tip forward and the ventral side upward. Pupae were injected with blue food colouring (McCormick) into the exuvium for 1–2 s by gently piercing the pupal case at the abdominal tip with the needle. During successful injections, no fluid was discharged from the pupa when the needle was removed, and the moulting fluid inside the exuvium was immediately stained. Pupae were washed in water three times to remove any excess dye. Following injections, 10 pupae were reared in social isolation to confirm the secretion of dyed droplets. For experiments, injected pupae were transferred to colonies with adult ants (Figs. 1f and  4c) or to colonies with adult ants and larvae (Figs. 3b and  4c) to track the distribution of the pupal social fluid.After spending 24 h with dye-injected pupae, adults were taken out of the colony, briefly immersed in 95% ethanol, and transferred to PBS. Digestive systems were dissected in cold PBS and mounted in DAKO mounting medium. Crop and stomach images (Fig. 1f, inset and Fig. 4c, inset) were acquired with a Revolve microscope (Echo). Larvae are translucent, and the presence of dye in the digestive system can be assayed without dissection. Whole-body images of larvae were acquired with a Leica Z16 APO microscope equipped with a Leica DFC450 camera and Leica Application Suite version 4.12.0 (Leica Microsystems). In the experiment on larval growth (Fig. 3c), larval length was measured from images using ImageJ32.Occluding pupaeTen pupae were placed on double-sided tape on a glass coverslip with the ventral side up. The area between the pupae was covered with laser-cut filter paper to prevent adults from sticking to the tape. The pupae were then placed in a 5 cm diameter Petri dish with a moist plaster of Paris floor. To block pupal secretion, the tip of the gaster was occluded with a drop of oil-paint (Uni Paint Markers PX-20), which has no discernible toxic effect7. Secreting pupae received a drop of the same paint on their head to control for putative differences resulting from the paint. Pupae were left in isolation for one day before adults were added to the assay chamber.Behavioural tracking of adult preference assayVideos were recorded using BFS-U3-50S5C-C: 5.0 MP, 35 FPS, Sony IMX264, Colour cameras (FLIR) and the Motif Video Recording System (Loopbio). To assess adult preference (Fig. 1g), physical contact of adults with pupae was manually annotated for the first 10 min after the first adult had encountered (physically contacted) a pupa.Protein profilingWe extracted 30 µl of pupal social fluid and whole-body wash samples with 75:25:0.2 acetonitrile: methanol: formic acid. Extracts were vortexed for 10 min, centrifuged at 16,000g and 4 °C for 10 min, dried in a SpeedVac, and stored at −80 °C until they were analysed by LC–MS/MS.Protein pellets were dissolved in 8 M urea, 50 mM ammonium bicarbonate, and 10 mM dithiothreitol, and disulfide bonds were reduced for 1 h at room temperature. Alkylation was performed by adding iodoacetamide to a final concentration of 20 mM and incubating for 1 h at room temperature in the dark. Samples were diluted using 50 mM ammonium bicarbonate until the concentration of urea had reached 3.5 M, and proteins were digested with endopeptidase LysC overnight at room temperature. Samples were further diluted to bring the urea concentration to 1.5 M before sequencing-grade modified trypsin was added. Digestion proceeded for 6 h at room temperature before being halted by acidification with TFA and samples were purified using in-house constructed C18 micropurification tips.LC–MS/MS analysis was performed using a Dionex3000 nanoflow HPLC and a Q-Exactive HF mass spectrometer (both Thermo Scientific). Solvent A was 0.1% formic acid in water and solvent B was 80% acetonitrile, 0.1% formic acid in water. Peptides were separated on a 90-minute linear gradient at 300 nl min−1 across a 75 µm × 100 mm fused-silica column packed with 3 µm Reprosil C18 material (Dr. Maisch). The mass spectrometer operated in positive ion Top20 DDA mode at resolution 60 k/30 k (MS1/MS2) and AGC targets were 3 × 106/2 × 105 (MS1/MS2).Raw files were searched through Proteome Discoverer v.1.4 (Thermo Scientific) and spectra were queried against the O. biroi proteome using MASCOT with a 1% FDR applied. Oxidation of M and acetylation of protein N termini were applied as a variable modification and carbamidomethylation of C was applied as a static modification. The average area of the three most abundant peptides for a matched protein33 was used to gauge protein amounts within and between samples.Functional annotation and gene ontology enrichmentTo supplement the current functional annotation of the O. biroi genome34, the full proteome for canonical transcripts was retrieved from UniProtKB (UniProt release 2020_04) in FASTA format. We then applied the EggNog-Mapper tool35,36 (http://eggnog-mapper.embl.de, emapper version 1.0.3-35-g63c274b, EggNogDB version 2) using standard parameters (m diamond -d none –tax_scope auto –go_evidence non-electronic –target_orthologs all –seed_ortholog_evalue 0.001 –seed_ortholog_score 60 –query-cover 20 –subject-cover 0) to produce an expanded annotation for all GO trees (Molecular Function, Biological Process, Cellular Components). The list of proteins identified in the pupal fluid was evaluated for functional enrichment in these GO terms, P-values were adjusted with an FDR cut-off of 0.05, and the network plots were visualized using the clusterProfiler package37.Metabolite profilingFor bulk polar metabolite profiling, we used 10 µl aliquots of pupal social fluid and whole-body wash (pooled samples). For the time-series metabolite profiling, 1 µl of pupal social fluid and whole-body wash was used. Samples were extracted in 180 µl cold LC–MS grade methanol containing 1 μM of uniformly labelled 15N- and 13C-amino acid internal standards (MSK-A2-1.2, Cambridge Isotope Laboratories) and consecutive addition of 390 µl LC–MS grade chloroform followed by 120 µl of LC–MS grade water.The samples were vortexed vigorously for 10 min followed by centrifugation (10 min at 16,000g and 4 °C). The upper polar metabolite-containing layer was collected, flash frozen and SpeedVac-dried. Dried extracts were stored at −80 °C until LC–MS analysis.LC–MS was conducted on a Q-Exactive benchtop Orbitrap mass spectrometer equipped with an Ion Max source and a HESI II probe, which was coupled to a Vanquish UPLC system (Thermo Fisher Scientific). External mass calibration was performed using the standard calibration mixture every three days.Dried polar samples were resuspended in 60 µl 50% acetonitrile, and 5 µl were injected into a ZIC-pHILIC 150 × 2.1 mm (5 µm particle size) column (EMD Millipore). Chromatographic separation was achieved using the following conditions: buffer A was 20 mM ammonium carbonate, 0.1% (v/v) ammonium hydroxide (adjusted to pH 9.3); buffer B was acetonitrile. The column oven and autosampler tray were held at 40 °C and 4 °C, respectively. The chromatographic gradient was run at a flow rate of 0.150 ml min−1 as follows: 0–22 min: linear gradient from 90% to 40% B; 22–24 min: held at 40% B; 24–24.1 min: returned to 90% B; 24.1 −30 min: held at 90% B. The mass spectrometer was operated in full-scan, polarity switching mode with the spray voltage set to 3.0 kV, the heated capillary held at 275 °C, and the HESI probe held at 250 °C. The sheath gas flow was set to 40 units, the auxiliary gas flow was set to 15 units. The MS data acquisition was performed in a range of 55–825 m/z, with the resolution set at 70,000, the AGC target at 10 × 106, and the maximum injection time at 80 ms. Relative quantification of metabolite abundances was performed using Skyline Daily v 20.1 (MacCoss Lab) with a 2 ppm mass tolerance and a pooled library of metabolite standards to confirm metabolite identity (via data-dependent acquisition). Metabolite levels were normalized by the mean signal of 8 heavy 13C,15N-labelled amino acid internal standards (technical normalization).The raw data were searched for a targeted list of ~230 polar metabolites and the corresponding peaks were integrated manually using Skyline Daily software. We were able to assign peaks to 107 compounds based on high mass accuracy ( More

Hawley, W. A. The biology of Aedes albopictus. J. Am. Mosq. Control Assoc. 1, 1–39 (1988).CAS 

Google Scholar 
Benedict, M. Q., Levine, R. S., Hawley, W. A. & Lounibos, L. P. Spread of the tiger: global risk of invasion by the mosquito Aedes albopictus. Vector-Borne Zoonotic Dis. 7, 76–85 (2007).Article 
PubMed 

Google Scholar 
Paupy, C., Delatte, H., Bagny, L., Corbel, V. & Fontenille, D. Aedes albopictus, an arbovirus vector: From the darkness to the light. Microb. Infect. 11, 1177–1185 (2009).Article 
CAS 

Google Scholar 
Delatte, H. et al. Blood-feeding behavior of Aedes albopictus, a vector of Chikungunya on La Réunion. Vector-Borne Zoonotic Dis. 10, 249–258 (2010).Article 
PubMed 

Google Scholar 
Pereira-dos-Santos, T., Roiz, D., Lourenço-de-Oliveira, R. & Paupy, C. A systematic review: Is Aedes albopictus an efficient bridge vector for zoonotic arboviruses? Pathogens 9, 266 (2020).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Gratz, N. Critical review of the vector status of Aedes albopictus. Med. Vet. Entomol. 18, 215–227 (2004).Article 
CAS 
PubMed 

Google Scholar 
Grard, G. et al. Zika virus in Gabon (Central Africa)—2007: A new threat from Aedes albopictus? PLoS Negl. Trop. Dis. 8, e2681 (2014).Article 
PubMed 
PubMed Central 

Google Scholar 
Lambrechts, L., Scott, T. W. & Gubler, D. J. Consequences of the expanding global distribution of Aedes albopictus for dengue virus transmission. PLoS Negl. Trop. Dis. 4, e646 (2010).Article 
PubMed 
PubMed Central 

Google Scholar 
Lounibos, L. P. & Kramer, L. D. Invasiveness of Aedes aegypti and Aedes albopictus and vectorial capacity for chikungunya virus. J. Infect. Dis. 214, S453–S458 (2016).Article 
PubMed 
PubMed Central 

Google Scholar 
European Centre for Disease Prevention and Control (ECDC). Vector Control with a Focus on Aedes aegypti and Aedes albopictus Mosquitoes: Literature Review and Analysis of Information (ECDC, Stockholm, Sweden, 2017).Tatem, A. J., Hay, S. I. & Rogers, D. J. Global traffic and disease vector dispersal. PNAS 103, 6242–6247 (2006).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Lowe, S., Browne, M., Boudjelas, S. & De Poorter, M. 100 of the World’s Worst Invasive Alien Species: A Selection From the Global Invasive Species Database, Vol. 12 (Invasive Species Specialist Group, 2000).Diagne, C. et al. High and rising economic costs of biological invasions worldwide. Nature 592, 571–576 (2021).Article 
CAS 
PubMed 

Google Scholar 
Hulme, P. E. Trade, transport and trouble: Managing invasive species pathways in an era of globalization. J. Appl. Ecol. 46, 10–18 (2009).Article 

Google Scholar 
Marini, F., Caputo, B., Pombi, M., Tarsitani, G. & Della-Torre, A. Study of Aedes albopictus dispersal in Rome, Italy, using sticky traps in mark–release–recapture experiments. Med. Vet. Entomol. 24, 361–368 (2010).Article 
CAS 
PubMed 

Google Scholar 
Bonizzoni, M., Gasperi, G., Chen, X. & James, A. A. The invasive mosquito species Aedes albopictus: current knowledge and future perspectives. Trends Parasitol. 29, 460–468 (2013).Article 
PubMed 
PubMed Central 

Google Scholar 
Collantes, F. et al. Review of ten-years presence of Aedes albopictus in Spain 2004–2014: Known distribution and public health concerns. Parasit Vectors 8, 1–11 (2015).Article 

Google Scholar 
Aranda, C., Eritja, R. & Roiz, D. First record and establishment of the mosquito Aedes albopictus in Spain. Med. Vet. Entomol. 20, 150–152 (2006).Article 
CAS 
PubMed 

Google Scholar 
Giménez, N. et al. Introduction of Aedes albopictus in Spain: A new challenge for public health. Gac. Sanit. 21, 25–28 (2007).Article 
PubMed 

Google Scholar 
European Centre for Disease Prevention and Control and European Food Safety Authority. Mosquito maps [internet]. Stockholm: ECDC. https://ecdc.europa.eu/en/disease-vectors/surveillance-and-disease-data/mosquito-maps (2022).Shigesada, N. & Kawasaki, K. Biological Invasions: Theory and Practice (Oxford University Press, 1997).
Google Scholar 
Puth, L. M. & Post, D. M. Studying invasion: Have we missed the boat? Ecol. Lett. 8, 715–721 (2005).Article 

Google Scholar 
Leung, B. et al. An ounce of prevention or a pound of cure: Bioeconomic risk analysis of invasive species. Proc. R Soc. Lond. Ser. B Biol. Sci. 269, 2407–2413 (2002).Article 

Google Scholar 
Lounibos, L. P. Invasions by insect vectors of human disease. Annu. Rev. Entomol. 47, 233–266 (2002).Article 
CAS 
PubMed 

Google Scholar 
Manni, M. et al. Genetic evidence for a worldwide chaotic dispersion pattern of the arbovirus vector, Aedes albopictus. PLoS Negl. Trop. Dis. 11, e0005332 (2017).Article 
PubMed 
PubMed Central 

Google Scholar 
Roiz, D. et al. Integrated Aedes management for the control of Aedes-borne diseases. PLoS Negl. Trop. Dis. 12, e0006845 (2018).Article 
PubMed 
PubMed Central 

Google Scholar 
Lühken, R. et al. Microsatellite typing of Aedes albopictus (Diptera: Culicidae) populations from Germany suggests regular introductions. Infect. Genet. Evol. 81, 104237 (2020).Article 
PubMed 

Google Scholar 
Battaglia, V. et al. The worldwide spread of the tiger mosquito as revealed by mitogenome haplogroup diversity. Front. Genet. 7, 208 (2016).Article 
PubMed 
PubMed Central 

Google Scholar 
Medley, K. A., Jenkins, D. G. & Hoffman, E. A. Human-aided and natural dispersal drive gene flow across the range of an invasive mosquito. Mol. Ecol. 24, 284–295 (2015).Article 
PubMed 

Google Scholar 
Eritja, R., Palmer, J. R., Roiz, D., Sanpera-Calbet, I. & Bartumeus, F. Direct evidence of adult Aedes albopictus dispersal by car. Sci. Rep. 7, 1–15 (2017).Article 
CAS 

Google Scholar 
Sherpa, S. et al. Unravelling the invasion history of the Asian tiger mosquito in Europe. Mol. Ecol. 28, 2360–2377 (2019).Article 
PubMed 

Google Scholar 
Swan, T. et al. A literature review of dispersal pathways of Aedes albopictus across different spatial scales: Implications for vector surveillance. Parasit Vectors 15, 1–13 (2022).Article 

Google Scholar 
Ballard, J. W. O. & Whitlock, M. C. The incomplete natural history of mitochondria. Mol. Ecol. 13, 729–744. https://doi.org/10.1046/j.1365-294X.2003.02063.x (2004).Article 
PubMed 

Google Scholar 
Toews, D. P. L. & Brelsford, A. The biogeography of mitochondrial and nuclear discordance in animals. Mol. Ecol. 21, 3907–3930. https://doi.org/10.1111/j.1365-294X.2012.05664.x (2012).Article 
CAS 
PubMed 

Google Scholar 
Hurst, G. D. & Jiggins, F. M. Problems with mitochondrial DNA as a marker in population, phylogeographic and phylogenetic studies: The effects of inherited symbionts. Proc. R. Soc. B: Biol. Sci. 272, 1525–1534 (2005).Article 
CAS 

Google Scholar 
Cariou, M., Duret, L. & Charlat, S. The global impact of Wolbachia on mitochondrial diversity and evolution. J. Evol. Biol. 30, 2204–2210 (2017).Article 
CAS 
PubMed 

Google Scholar 
Zug, R. & Hammerstein, P. Still a host of hosts for Wolbachia: analysis of recent data suggests that 40% of terrestrial arthropod species are infected. PLoS ONE 7, e38544 (2012).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Weinert, L. A., Araujo-Jnr, E. V., Ahmed, M. Z. & Welch, J. J. The incidence of bacterial endosymbionts in terrestrial arthropods. Proc. R. Soc. B: Biol. Sci. 282, 20150249 (2015).Article 

Google Scholar 
Goubert, C., Minard, G., Vieira, C. & Boulesteix, M. Population genetics of the Asian tiger mosquito Aedes albopictus, an invasive vector of human diseases. Heredity 117, 125–134 (2016).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Western, D. Human-modified ecosystems and future evolution. PNAS 98, 5458–5465 (2001).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Pech-May, A. et al. Population genetics and ecological niche of invasive Aedes albopictus in Mexico. Acta Trop. 157, 30–41 (2016).Article 
PubMed 

Google Scholar 
Vargo, E. L. et al. Hierarchical genetic analysis of German cockroach (Blattella germanica) populations from within buildings to across continents. PLoS ONE 9, e102321 (2014).Article 
PubMed 
PubMed Central 

Google Scholar 
von Beeren, C., Stoeckle, M. Y., Xia, J., Burke, G. & Kronauer, D. J. Interbreeding among deeply divergent mitochondrial lineages in the American cockroach (Periplaneta americana). Sci. Rep. 5, 1–7 (2015).
Google Scholar 
Tseng, S.-P. et al. Genetic diversity and Wolbachia infection patterns in a globally distributed invasive ant. Front. Genet. 10, 838 (2019).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Wesson, D. M., Porter, C. H. & Collins, F. H. Sequence and secondary structure comparisons of ITS rDNA in mosquitoes (Diptera: Culicidae). Mol. Phylogen. Evol. 1, 253–269 (1992).Article 
CAS 

Google Scholar 
Mishra, S., Sharma, G., Das, M. K., Pande, V. & Singh, O. P. Intragenomic sequence variations in the second internal transcribed spacer (ITS2) ribosomal DNA of the malaria vector Anopheles stephensi. PLoS ONE 16, e0253173 (2021).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Artigas, P. et al. Aedes albopictus diversity and relationships in south-western Europe and Brazil by rDNA/mtDNA and phenotypic analyses: ITS-2, a useful marker for spread studies. Parasit Vectors 14, 1–23 (2021).Article 

Google Scholar 
Armbruster, P. et al. Infection of New-and Old-World Aedes albopictus (Diptera: Culicidae) by the intracellular parasite Wolbachia: implications for host mitochondrial DNA evolution. J. Med. Entomol. 40, 356–360 (2003).Article 
PubMed 

Google Scholar 
Maia, R., Scarpassa, V. M., Maciel-Litaiff, L. & Tadei, W. P. Reduced levels of genetic variation in Aedes albopictus (Diptera: Culicidae) from Manaus, Amazonas State, Brazil, based on analysis of the mitochondrial DNA ND5 gene. Gen. Mol. Res. 2000, 998–1007 (2009).Article 

Google Scholar 
Birungi, J. & Munstermann, L. E. Genetic structure of Aedes albopictus (Diptera: Culicidae) populations based on mitochondrial ND5 sequences: Evidence for an independent invasion into Brazil and United States. Ann. Entomol. Soc. Am. 95, 125–132 (2002).Article 
CAS 

Google Scholar 
Kambhampati, S. & Rai, K. S. Mitochondrial DNA variation within and among populations of the mosquito Aedes albopictus. Genome 34, 288–292 (1991).Article 
CAS 
PubMed 

Google Scholar 
Werren, J. H., Baldo, L. & Clark, M. E. Wolbachia: master manipulators of invertebrate biology. Nat. Rev. Microbiol. 6, 741–751 (2008).Article 
CAS 
PubMed 

Google Scholar 
Wiwatanaratanabutr, I. Geographic distribution of wolbachial infections in mosquitoes from Thailand. J. Invertebr. Pathol. 114, 337–340 (2013).Article 
PubMed 

Google Scholar 
Carvajal, T. M., Hashimoto, K., Harnandika, R. K., Amalin, D. M. & Watanabe, K. Detection of Wolbachia in field-collected Aedes aegypti mosquitoes in metropolitan Manila, Philippines. Parasit. Vectors 12, 1–9 (2019).Article 

Google Scholar 
Atyame, C. M., Delsuc, F., Pasteur, N., Weill, M. & Duron, O. Diversification of Wolbachia endosymbiont in the Culex pipiens mosquito. Mol. Biol. Evol. 28, 2761–2772 (2011).Article 
CAS 
PubMed 

Google Scholar 
Damiani, C. et al. Wolbachia in Aedes koreicus: Rare detections and possible implications. Insects 13, 216 (2022).Article 
PubMed 
PubMed Central 

Google Scholar 
Jiggins, F. M. Male-killing Wolbachia and mitochondrial DNA: Selective sweeps, hybrid introgression and parasite population dynamics. Genetics 164, 5–12 (2003).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Schuler, H. et al. The hitchhiker’s guide to Europe: The infection dynamics of an ongoing Wolbachia invasion and mitochondrial selective sweep in Rhagoletis cerasi. Mol. Ecol. 25, 1595–1609 (2016).Article 
PubMed 
PubMed Central 

Google Scholar 
Ross, P. A., Ritchie, S. A., Axford, J. K. & Hoffmann, A. A. Loss of cytoplasmic incompatibility in Wolbachia-infected Aedes aegypti under field conditions. PLoS Negl. Trop. Dis. 13, e0007357 (2019).Article 
PubMed 
PubMed Central 

Google Scholar 
Avise, J. C. Phylogeography: The history and formation of species (Harvard University Press, 2000).Book 

Google Scholar 
Rokas, A., Atkinson, R. J., Brown, G. S., West, S. A. & Stone, G. N. Understanding patterns of genetic diversity in the oak gallwasp Biorhiza pallida: Demographic history or a Wolbachia selective sweep? Heredity 87, 294–304 (2001).Article 
CAS 
PubMed 

Google Scholar 
Porretta, D., Mastrantonio, V., Bellini, R., Somboon, P. & Urbanelli, S. Glacial history of a modern invader: Phylogeography and species distribution modelling of the Asian tiger mosquito Aedes albopictus. PLoS ONE 7, e44515. https://doi.org/10.1371/journal.pone.0044515 (2012).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Motoki, M. T. et al. Population genetics of Aedes albopictus (Diptera: Culicidae) in its native range in Lao People’s Democratic Republic. Parasit. Vectors 12, 1–12 (2019).Article 
CAS 

Google Scholar 
Zhong, D. et al. Genetic analysis of invasive Aedes albopictus populations in Los Angeles County, California and its potential public health impact. PLoS ONE 8, e68586 (2013).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Usmani-Brown, S., Cohnstaedt, L. & Munstermann, L. E. Population genetics of Aedes albopictus (Diptera: Culicidae) invading populations, using mitochondrial nicotinamide adenine dinucleotide dehydrogenase subunit 5 sequences. Ann. Entomol. Soc. Am. 102, 144–150 (2009).Article 
CAS 
PubMed 

Google Scholar 
Mousson, L. et al. Phylogeography of Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) based on mitochondrial DNA variations. Genet. Res. 86, 1–11 (2005).Article 
CAS 
PubMed 

Google Scholar 
Bazin, E., Glémin, S. & Galtier, N. Population size does not influence mitochondrial genetic diversity in animals. Science 312, 570–572. https://doi.org/10.1126/science.1122033 (2006).Article 
CAS 
PubMed 

Google Scholar 
Dowling, D. K., Friberg, U. & Lindell, J. Evolutionary implications of non-neutral mitochondrial genetic variation. Ecol. Evol. 23, 546–554 (2008).Article 

Google Scholar 
Montero-Pau, J., Gómez, A. & Muñoz, J. Application of an inexpensive and high-throughput genomic DNA extraction method for the molecular ecology of zooplanktonic diapausing eggs. Limnol. Oceanogr. Methods 6, 218–222 (2008).Article 
CAS 

Google Scholar 
Porter, C. H. & Collins, F. H. Species-diagnostic differences in a ribosomal DNA internal transcribed spacer from the sibling species Anopheles freeborni and Anopheles hermsi (Diptera: Culicidae). Am. J. Trop. Med. 45, 271–279 (1991).Article 
CAS 

Google Scholar 
Folmer, O., Black, M., Hoeh, W., Lutz, R. & Vrijenhoek, R. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol. Mar. Biol. Biotechnol. 3, 294–299 (1994).CAS 
PubMed 

Google Scholar 
Prosser, S., Martínez-Arce, A. & Elías-Gutiérrez, M. A new set of primers for COI amplification from freshwater microcrustaceans. Mol. Ecol. Resour. 13, 1151–1155 (2013).CAS 
PubMed 

Google Scholar 
Ivanova, N. V., Zemlak, T. S., Hanner, R. H. & Hebert, P. D. Universal primer cocktails for fish DNA barcoding. Mol. Ecol. Notes 7, 544–548 (2007).Article 
CAS 

Google Scholar 
Kumar, S., Stecher, G. & Tamura, K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 33, 1870–1874 (2016).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Zhou, W., Rousset, F. & O’Neill, S. Phylogeny and PCR–based classification of Wolbachia strains using wsp gene sequences. Proc. R Soc. Lond. Ser. B Biol. Sci. 265, 509–515 (1998).Article 
CAS 

Google Scholar 
Braig, H. R., Zhou, W., Dobson, S. L. & O’Neill, S. L. Cloning and characterization of a gene encoding the major surface protein of the bacterial endosymbiont Wolbachia pipientis. J. Bacteriol. 180, 2373–2378 (1998).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Hu, Y. et al. Identification and molecular characterization of Wolbachia strains in natural populations of Aedes albopictus in China. Parasit. Vectors 13, 1–14 (2020).
Google Scholar 
Heddi, A., Grenier, A.-M., Khatchadourian, C., Charles, H. & Nardon, P. Four intracellular genomes direct weevil biology: Nuclear, mitochondrial, principal endosymbiont, and Wolbachia. PNAS 96, 6814–6819 (1999).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Rozas, J. et al. DnaSP 6: DNA sequence polymorphism analysis of large data sets. Mol. Biol. Evol. 34, 3299–3302 (2017).Article 
CAS 
PubMed 

Google Scholar 
Salzburger, W., Ewing, G. B. & Von Haeseler, A. The performance of phylogenetic algorithms in estimating haplotype genealogies with migration. Mol. Ecol. 20, 1952–1963 (2011).Article 
PubMed 

Google Scholar 
Stamatakis, A. RAxML-VI-HPC: Maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics 22, 2688–2690 (2006).Article 
CAS 
PubMed 

Google Scholar 
Darriba, D., Taboada, G. L., Doallo, R. & Posada, D. jModelTest 2: More models, new heuristics and parallel computing. Nat. Methods 9, 772. https://doi.org/10.1038/nmeth.2109 (2012).Article 
CAS 
PubMed 
PubMed Central 

Google Scholar 
Gower, J. C. Some distance properties of latent root and vector methods used in multivariate analysis. Biometrika 53, 325–338 (1966).Article 
MathSciNet 
MATH 

Google Scholar 
R Core Team. R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing. http://www.R-project.org/. (2021).Hijmans, R. J., Williams, E., Vennes, C. & Hijmans, M. R. J. Package ‘geosphere’. Spher. Trigon. 1, 5 (2017).
Google Scholar 
Palmer, J. R. et al. Citizen science provides a reliable and scalable tool to track disease-carrying mosquitoes. Nat. Commun. 8, 1–13 (2017).Article 

Google Scholar 
Mantel, N. & Valand, R. S. A technique of nonparametric multivariate analysis. Biometrics 1970, 547–558 (1970).Article 

Google Scholar 
Goslee, S. C. & Urban, D. L. The ecodist package for dissimilarity-based analysis of ecological data. J. Stat. Softw. 22, 1–19 (2007).Article 

Google Scholar 
Stewart, C. Zero-inflated beta distribution for modeling the proportions in quantitative fatty acid signature analysis. J. Appl. Stat. 40, 985–992 (2013).Article 
MathSciNet 
MATH 

Google Scholar 
Figueroa-Zúñiga, J. I., Arellano-Valle, R. B. & Ferrari, S. L. Mixed beta regression: A Bayesian perspective. Comput. Stat. Data Anal. 61, 137–147 (2013).Article 
MathSciNet 
MATH 

Google Scholar 
Branscum, A. J., Johnson, W. O. & Thurmond, M. C. Bayesian beta regression: Applications to household expenditure data and genetic distance between foot-and-mouth disease viruses. Aust. N. Z. J. Stat. 49, 287–301 (2007).Article 
MathSciNet 
MATH 

Google Scholar 
Ospina, R. & Ferrari, S. L. Inflated beta distributions. Stat. Pap. 51, 111–126 (2010).Article 
MathSciNet 
MATH 

Google Scholar 
Chung, H. & Beretvas, S. N. The impact of ignoring multiple membership data structures in multilevel models. Br. J. Math. Stat. Psychol. 65, 185–200 (2012).Article 
MathSciNet 
PubMed 
MATH 

Google Scholar  More

arising from Y. Wang et al. Nature https://doi.org/10.1038/s41586-021-04016-x (2021)A unique challenge for environmental DNA (eDNA)-based palaeoecological reconstructions and extinction estimates is that organisms can contribute DNA to sediments long after their death. Recently, Wang et al.1 discovered mammoth eDNA in sediments that are between approximately 4.6 and 7 thousand years (kyr) younger than the most recent mammoth fossils in North America and Eurasia, which they interpreted as mammoths surviving on both continents into the Middle Holocene epoch. Here we present an alternative explanation for these offsets: the slow decomposition of mammoth tissues on cold Arctic landscapes is responsible for the release of DNA into sediments for thousands of years after mammoths went extinct. eDNA records are important palaeobiological archives, but the mixing of undatable DNA from long-dead organisms into younger sediments complicates the interpretation of eDNA, particularly from cold and high-latitude systems.All animal tissues, including faeces, contribute DNA to eDNA records2, but the durations across which tissues can contribute genetic information must vary depending on tissue type and local rates of destruction and decomposition. On high-latitude landscapes, soft tissues and skeletal remains of large mammals may persist, unburied, for millennia3,4,5. For example, unburied antlers of caribou (Rangifer tarandus) from Svalbard (Norway) and Ellesmere Island (Canada) have been dated3,4 to between 1 and 2 cal kyr bp (calibrated kyr before present). Elephant seal (Mirounga leonina) remains near the Antarctic coastline5,6 can persist for more than 5,000 years. This is in contrast to bones in warmer settings, which persist for only centuries or decades7,8. Because bones are particularly resistant to decay, quantifying how their persistence changes across environments enables us to constrain the durations that dead individuals generally contribute to eDNA archives. To do this, we consolidated data on the oldest radiocarbon-dated surface-collected bones from different ecosystems. We included bones that we are reasonably confident persisted without being completely buried (‘never buried’), and bones for which exhumation cannot be confidently excluded (‘potentially never buried’). Pairing bone persistence with mean annual temperatures (MAT) from their sample localities, we find a strong link between the local temperature and the logged duration of bone persistence (Fig. 1, never buried bones: R2 = 0.94, P  More

Olszewska-Guizzo, A., Fogel, A., Benjumea, D. & Tahsin, N. Sustainable Policies and Practices in Energy, Environment and Health Research 223–243 (Springer, 2022).Book 

Google Scholar 
Gascon, M. et al. Mental health benefits of long-term exposure to residential green and blue spaces: A systematic review. Int. J. Environ. Res. Public Health 12, 4354–4379. https://doi.org/10.3390/ijerph120404354 (2015).Article 
PubMed 
PubMed Central 

Google Scholar 
Houlden, V., Weich, S., Porto-de-Albuquerque, J., Jarvis, S. & Rees, K. The relationship between greenspace and the mental wellbeing of adults: A systematic review. PLoS ONE 13, 3000 (2018).Article 

Google Scholar 
Hung, S.-H. & Chang, C.-Y. Health benefits of evidence-based biophilic-designed environments: A review. J. People Plants Env. 24, 1–16 (2021).Article 

Google Scholar 
Berman, M. G., Jonides, J. & Kaplan, S. The cognitive benefits of interacting with nature. Psychol. Sci. 19, 1207–1212. https://doi.org/10.1111/j.1467-9280.2008.02225.x (2008).Article 
PubMed 

Google Scholar 
Kaplan, S. Meditation, restoration, and the management of mental fatigue. Environ. Behav. 33, 480–506. https://doi.org/10.1177/00139160121973106 (2001).Article 

Google Scholar 
Ulrich, R. S. et al. Stress recovery during exposure to natural and urban environments. J. Environ. Psychol. 11, 201–230 (1991).Article 

Google Scholar 
Kellert, S. R. & Wilson, E. O. The Biophilia Hypothesis (Island Press, 1993).
Google Scholar 
Stack, K. & Shultis, J. Implications of attention restoration theory for leisure planners and managers. Leisure/Loisir 37, 1–16 (2013).Article 

Google Scholar 
Steel, Z. et al. The global prevalence of common mental disorders: A systematic review and meta-analysis 1980–2013. Int. J. Epidemiol. 43, 476–493. https://doi.org/10.1093/ije/dyu038 (2014).Article 
PubMed 
PubMed Central 

Google Scholar 
Mueller, D. P. The current status of urban-rural differences in psychiatric disorder. An emerging trend for depression. J. Nerv. Ment. Dis. 169, 18–27 (1981).Article 
CAS 
PubMed 

Google Scholar 
Peen, J., Schoevers, R. A., Beekman, A. T. & Dekker, J. The current status of urban-rural differences in psychiatric disorders. Acta Psychiatr. Scand. 121, 84–93. https://doi.org/10.1111/j.1600-0447.2009.01438.x (2010).Article 
CAS 
PubMed 

Google Scholar 
Taylor, L. & Hochuli, D. F. Defining greenspace: Multiple uses across multiple disciplines. Landsc. Urban Plan. 158, 25–38 (2017).Article 

Google Scholar 
en K Staats, H. Restorative Environments The Oxford Handbook of Environmental and Conservation Psychology 445th edn. (Oxford University Press, 2012).
Google Scholar 
Wood, L., Hooper, P., Foster, S. & Bull, F. Public green spaces and positive mental health–investigating the relationship between access, quantity and types of parks and mental wellbeing. Health Place 48, 63–71 (2017).Article 
PubMed 

Google Scholar 
Tsunetsugu, Y. et al. Physiological and psychological effects of viewing urban forest landscapes assessed by multiple measurements. Landsc. Urban Plan. 113, 90–93 (2013).Article 

Google Scholar 
Gidlow, C. J. et al. Where to put your best foot forward: Psycho-physiological responses to walking in natural and urban environments. J. Environ. Psychol. 45, 22–29 (2016).Article 

Google Scholar 
Lee, J. Experimental study on the health benefits of garden landscape. Int. J. Environ. Res. Public Health 14, 829 (2017).Article 
PubMed 
PubMed Central 

Google Scholar 
Fuller, R. A., Irvine, K. N., Devine-Wright, P., Warren, P. H. & Gaston, K. J. Psychological benefits of greenspace increase with biodiversity. Biol. Lett. 3, 390–394 (2007).Article 
PubMed 
PubMed Central 

Google Scholar 
Thompson, C. W., Aspinall, P. & Bell, S. Innovative Approaches to Researching Landscape and Health: Open Space: People Space 2 (Routledge, 2010).Book 

Google Scholar 
Tsutsumi, M., Nogaki, H., Shimizu, Y., Stone, T. E. & Kobayashi, T. Individual reactions to viewing preferred video representations of the natural environment: A comparison of mental and physical reactions. Jpn. J. Nurs. Sci. 14, 3–12 (2017).Article 
PubMed 

Google Scholar 
Grazuleviciene, R. et al. Tracking restoration of park and urban street settings in coronary artery disease patients. Int. J. Environ. Res. Public Health 13, 550 (2016).Article 
PubMed 
PubMed Central 

Google Scholar 
Bostancı, S. H. In New Approaches to Spatial Planning and Design (ed Murat Özyavuz) Ch. 32, 435–451 (Peter Lang, 2019).Daniel, T. C. Measuring Landscape Esthetics: The Scenic Beauty Estimation Method, vol. 167 (Department of Agriculture, Forest Service, Rocky Mountain Forest and Range…, 1976).Bacon, W. R. In (eds Elsner G. H. et al) Technical Coordinators. Proceedings of our national landscape: A conference on applied techniques for analysis and management of the visual resource [Incline Village, Nev., April 23–25, 1979]. Gen. Tech. Rep. PSW-GTR-35. Berkeley, CA. Pacific Southwest Forest and Range Exp. Stn., Forest Service, US Department of Agriculture 660–665 (1979).Gavrilidis, A. A., Ciocănea, C. M., Niţă, M. R., Onose, D. A. & Năstase, I. I. Urban landscape quality index—planning tool for evaluating urban landscapes and improving the quality of life. Procedia Environ. Sci. 32, 155–167. https://doi.org/10.1016/j.proenv.2016.03.020 (2016).Article 

Google Scholar 
Knobel, P. et al. Development of the urban green space quality assessment tool (RECITAL). Urban For. Urban Green. 57, 126895 (2021).Article 

Google Scholar 
Bacon, W. R. & Dell, J. National Forest Landscape Management (Forest Service, US Department of Agriculture, 1973).Kaplan, R., Kaplan, S. & Ryan, R. With People in Mind: Design and Management of Everyday Nature (Island Press, 1998).
Google Scholar 
Smardon, R., Palmer, J. & Felleman, J. P. Foundations for Visual Project Analysis (Wiley, 1986).
Google Scholar 
Jung, C. G. Man and His Symbols Garden City (Doubleday and Co, 1964).
Google Scholar 
Olszewska, A., Marques, P. F., Ryan, R. L. & Barbosa, F. What makes a landscape contemplative?. Env. Plan. B Urban Anal. City Sci. 45, 7–25. https://doi.org/10.1177/0265813516660716 (2016).Article 

Google Scholar 
Tarkka, I. M. & Hallett, M. Cortical topography of premotor and motor potentials preceding self-paced, voluntary movement of dominant and non-dominant hands. Electroencephalogr. Clin. Neurophysiol. 75, 36–43 (1990).Article 
CAS 
PubMed 

Google Scholar 
Olszewska-Guizzo, A., Paiva, T. O. & Barbosa, F. Effects of 3D contemplative landscape videos on brain activity in a passive exposure EEG experiment. Front. Psychiatry 9, 317. https://doi.org/10.3389/fpsyt.2018.00317 (2018).Article 
PubMed 
PubMed Central 

Google Scholar 
Bradley, M. M. & Lang, P. J. Measuring emotion: The self-assessment manikin and the semantic differential. J. Behav. Ther. Exp. Psychiatry 25, 49–59. https://doi.org/10.1016/0005-7916(94)90063-9 (1994).Article 
CAS 
PubMed 

Google Scholar 
Beck, A. T., Steer, R. A. & Brown, G. K. Beck depression inventory-II. San Antonio 78, 490–498 (1996).
Google Scholar 
Ferree, T. C., Luu, P., Russell, G. S. & Tucker, D. M. Scalp electrode impedance, infection risk, and EEG data quality. Clin. Neurophysiol. 112, 536–544. https://doi.org/10.1016/S1388-2457(00)00533-2 (2001).Article 
CAS 
PubMed 

Google Scholar 
Stroganova, T. A. & Orekhova, E. V. EEG and infant states. Infant EEG Event-Relat. Potentials 251, 280 (2007).
Google Scholar 
Cacioppo, J. T., Tassinary, L. G. & Berntson, G. Handbook of Psychophysiology (Cambridge University Press, 2007).
Google Scholar 
Ulrich, R. S. Natural versus urban scenes: Some psychophysiological effects. Environ. Behav. 13, 523–556 (1981).Article 

Google Scholar 
Choi, Y., Kim, M. & Chun, C. Measurement of occupants’ stress based on electroencephalograms (EEG) in twelve combined environments. Build. Environ. 88, 65–72 (2015).Article 

Google Scholar 
Gorji, M. A. H., Davanloo, A. A. & Heidarigorji, A. M. The efficacy of relaxation training on stress, anxiety, and pain perception in hemodialysis patients. Indian J. Nephrol. 24, 356 (2014).Article 

Google Scholar 
Cahn, B. R. & Polich, J. Meditation states and traits: EEG, ERP, and neuroimaging studies. Psychol. Bull. 132, 180 (2006).Article 
PubMed 

Google Scholar 
Gruzelier, J. A theory of alpha/theta neurofeedback, creative performance enhancement, long distance functional connectivity and psychological integration. Cogn. Process. 10, 101–109 (2009).Article 

Google Scholar 
Vecchiato, G. et al. Neurophysiological correlates of embodiment and motivational factors during the perception of virtual architectural environments. Cogn. Process. 16, 425–429 (2015).Article 
PubMed 

Google Scholar 
Lagopoulos, J. et al. Increased theta and alpha EEG activity during nondirective meditation. J. Altern. Complement. Med. 15, 1187–1192 (2009).Article 
PubMed 

Google Scholar 
Wascher, E. et al. Frontal theta activity reflects distinct aspects of mental fatigue. Biol. Psychol. 96, 57–65 (2014).Article 
PubMed 

Google Scholar 
Kabat-Zinn, J. Mindfulness. Mindfulness 6, 1481–1483 (2015).Article 

Google Scholar 
McGarrigle, T. & Walsh, C. A. Mindfulness, self-care, and wellness in social work: Effects of contemplative training. J. Relig. Spiritual. Soc. Work Soc. Thought 30, 212–233 (2011).
Google Scholar 
Grossman, P., Niemann, L., Schmidt, S. & Walach, H. Mindfulness-based stress reduction and health benefits: A meta-analysis. J. Psychosom. Res. 57, 35–43 (2004).Article 
PubMed 

Google Scholar 
Bailey, A. W., Allen, G., Herndon, J. & Demastus, C. Cognitive benefits of walking in natural versus built environments. World Leisure J. 60, 293–305 (2018).Article 

Google Scholar 
Qin, J., Zhou, X., Sun, C., Leng, H. & Lian, Z. Influence of green spaces on environmental satisfaction and physiological status of urban residents. Urban For. Urban Green. 12, 490–497 (2013).Article 

Google Scholar 
Kolb, B. & Whishaw, I. Q. Fundamentals of Human Neuropsychology (Freeman, 1990).
Google Scholar 
Milner, B. Visual recognition and recall after right temporal-lobe excision in man. Neuropsychologia 6, 191–209 (1968).Article 

Google Scholar 
Corbetta, M. & Shulman, G. L. Control of goal-directed and stimulus-driven attention in the brain. Nat. Rev. Neurosci. 3, 201–215. https://doi.org/10.1038/nrn755 (2002).Article 
CAS 
PubMed 

Google Scholar 
Chang, C.-Y. & Chen, P.-K. Human response to window views and indoor plants in the workplace. HortScience 40, 1354–1359 (2005).Article 

Google Scholar 
Herzog, T. R., Black, A. M., Fountaine, K. A. & Knotts, D. J. Reflection and attentional recovery as distinctive benefits of restorative environments. J. Environ. Psychol. 17, 165–170 (1997).Article 

Google Scholar 
Baehr, E., Rosenfeld, J. P. & Baehr, R. Clinical use of an alpha asymmetry neurofeedback protocol in the treatment of mood disorders: Follow-up study one to five years post therapy. J. Neurother. 4, 11–18 (2001).Article 

Google Scholar 
Sia, A. et al. Nature-based activities improve the well-being of older adults. Sci. Rep. 10, 1–8 (2020).Article 

Google Scholar 
Olszewska-Guizzo, A., Sia, A., Fogel, A. & Ho, R. Can exposure to certain urban green spaces trigger frontal alpha asymmetry in the brain?—Preliminary findings from a passive task EEG study. Int. J. Environ. Res. Public Health 17, 394 (2020).Article 
PubMed 
PubMed Central 

Google Scholar 
Olszewska-Guizzo, A. et al. Therapeutic garden with contemplative features induces desirable changes in mood and B rain activity in depressed adults. Front. Psychiatry https://doi.org/10.3389/fpsyt.2022.757056 (2021).Article 

Google Scholar 
Tan, S. B., Vignesh, L. N. & Donald, L. Public Housing in Singapore: Examining Fundamental Shifts (Lee Kuan Yew School of Public Policy, National University of Singapore, 2014).Tan, P. Y. Nature, Place & People: Forging Connections Through Neighbourhood Landscape Design (World Scientific Publishing Co., 2018).Book 

Google Scholar 
Peirce, J. et al. PsychoPy2: Experiments in behavior made easy. Behav. Res. Methods 51, 195–203. https://doi.org/10.3758/s13428-018-01193-y (2019).Article 
PubMed 
PubMed Central 

Google Scholar 
Edwards, A. L. Balanced Latin-square designs in psychological research. Am. J. Psychol. 64, 598–603 (1951).Article 
CAS 
PubMed 

Google Scholar 
Korpela, K. M., Ylén, M., Tyrväinen, L. & Silvennoinen, H. Determinants of restorative experiences in everyday favorite places. Health Place 14, 636–652 (2008).Article 
PubMed 

Google Scholar 
Ojala, A., Korpela, K., Tyrväinen, L., Tiittanen, P. & Lanki, T. Restorative effects of urban green environments and the role of urban-nature orientedness and noise sensitivity: A field experiment. Health Place 55, 59–70 (2019).Article 
PubMed 

Google Scholar 
Tyrväinen, L. et al. The influence of urban green environments on stress relief measures: A field experiment. J. Environ. Psychol. 38, 1–9 (2014).Article 

Google Scholar 
Herzog, T. R. & Barnes, G. J. Tranquility and preference revisited. J. Environ. Psychol. 19, 171–181 (1999).Article 

Google Scholar 
Neale, C. et al. The impact of walking in different urban environments on brain activity in older people. Cities Health 4, 94–106. https://doi.org/10.1080/23748834.2019.1619893 (2020).Article 

Google Scholar 
Kaplan, R. & Kaplan, S. The Experience of Nature: A Psychological Perspective (CUP Archive, 1989).
Google Scholar 
Treib, M. In Contemporary Landscapes of Contemplation (ed Rebecca Krinke) 27–49 (Routledge, 2005).Appleton, J. The Experience of Landscape (Wiley Chichester, 1996).
Google Scholar 
Grahn, P., Ottosson, J. & Uvnäs-Moberg, K. The oxytocinergic system as a mediator of anti-stress and instorative effects induced by nature: The calm and connection theory. Front. Psychol. 2021, 12 (2021).
Google Scholar 
Hartig, T., Mang, M. & Evans, G. W. Restorative effects of natural environment experiences. Environ. Behav. 23, 3–26. https://doi.org/10.1177/0013916591231001 (1991).Article 

Google Scholar 
Stamps Iii, A. E. Use of photographs to simulate environments: A meta-analysis. Percept. Mot. Skills 71, 907–913 (1990).Article 

Google Scholar 
Menardo, E., Brondino, M., Hall, R. & Pasini, M. Restorativeness in natural and urban environments: A meta-analysis. Psychol. Rep. 124, 417–437 (2021).Article 
PubMed 

Google Scholar  More

Kirilova, E. P., Cremer, H., Heiri, O. & Lotter, A. F. Eutrophication of moderately deep Dutch lakes during the past century: Flaws in the expectations of water management? Hydrobiologia 637, 157–171 (2010).Article 
CAS 

Google Scholar 
Scharf, B. & Viehberg, F. A. Living Ostracoda (Crustacea) from the town moat of Bremen, Germany. Crustaceana 87(8–9), 1124–1135 (2014).Article 

Google Scholar 
Rees, S. E. The historical and cultural importance of ponds and small lakes in Wales, UK. Aquat. Conserv. 7(2), 133–139 (1997).Article 

Google Scholar 
Brown, A. et al. The ecological impact of conquest and colonisation on a medieval frontier landscape: Combined palynological and geochemical analysis of lake sediments from Radzyń Chełmiński, northern Poland. Geoarchaeology 30, 511–527 (2015).Article 

Google Scholar 
Kittel, P. et al. The palaeoecological development of the Late Medieval moat—Multiproxy research at Rozprza Central Poland. Quat. Int. 482, 131–156 (2018).Article 

Google Scholar 
Hildebrandt-Radke, I. Geoarchaeological aspects in the studies of prehistoric and early historic settlement complexes. In Studia interdyscyplinarne nad środowiskiem i kulturą w Polsce. Tom 1. Środowisko-Człowiek-Cywilizacja (eds Makohonienko, M. et al.) 57–70 (Bogucki Wyd Naukowe, 2007).
Google Scholar 
Łyskowski, M. & Wardas-Lasoń, M. Georadar investigations and geochemical analysis in contemporary archaeological studies. Geol. Geophys. Environ. 38(3), 307–315 (2012).Article 

Google Scholar 
Korhola, A. & Rautio, M. Cladocera and other branchiopod crustaceans. In Tracking Environmental Change Using Lake Sediments, Vol. 4: Zoological Indicators (eds Smol, J. P. et al.) 5–41 (Kluwer Academic Publishers, 2001).Chapter 

Google Scholar 
Birks, H. H. Plant macrofossils. In Tracking Environmental Change Using Lake Sediments, 3: Terrestrial, Algal, and Siliceous Indicators (eds Smol, J. P. et al.) 49–74 (Kluwer Academic Publishers, 2001).
Google Scholar 
Battarbee, R. W. Diatom analysis. In Handbook of Holocene Palaeoecology and Palaeohydrology (ed. Berglund, B. E.) 527–570 (Wiley, 1986).
Google Scholar 
Luoto, T. P., Nevalainen, L., Kultti, S. & Sarmaja-Korjonen, K. An evaluation of the influence of water depth and river inflow on quantitative Cladocera-based temperature and lake level inferences in a shallow boreal lake. Hydrobiologia 676, 143–154 (2011).Article 
CAS 

Google Scholar 
Luoto, T. P. Intra-lake patterns of aquatic insect and mite remains. J. Paleolimnol. 47, 141–157 (2012).Article 

Google Scholar 
Hann, B. J. Methods in Quaternary ecology. Cladocera. Geosci. Canada 16, 17–26 (1989).
Google Scholar 
Dimbleby, G. W. The Palynology of Archaeological Sites (Academic Press. Inc., 1985).
Google Scholar 
Edwards, K. J. Using space in cultural palynology: The value of the off-site pollen record. In Modelling Ecological Change: Perspectives from Neoecology, Palaeoecology and Environmental Archaeology (eds Harris, D. R. & Thomas, K. D.) 61–74 (Routledge Taylor & Francis Group, 2016).
Google Scholar 
Kittel, P., Sikora, J. & Wroniecki, P. A Late Medieval motte-and-bailey settlement in a lowland river valley landscape of central Poland. Geoarchaeology 33(5), 558–578 (2018).Article 

Google Scholar 
Antczak-Orlewska, O. et al. The environmental history of the oxbow in the Luciąża River valley—Study on the specific microclimate during Allerød and Younger Dryas in central Poland. Quat. Int. https://doi.org/10.1016/j.quaint.2021.08.011 (2021).Article 

Google Scholar 
Dearing, J. A. Core correlation and total sediment influx. In Handbook of Holocene Palaeoecology and Palaeohydrology (ed. Berglund, B. E.) 247–270 (Wiley, 1986).
Google Scholar 
O’Brien, C. et al. A sediment-based multiproxy palaeoecological approach to the environmental archaeology of lake dwellings (crannogs), central Ireland. Holocene 15, 707–719 (2005).Article 

Google Scholar 
Ruiz, Z., Brown, A. G. & Langdon, P. G. The potential of chironomid (Insecta: Diptera) larvae in archaeological investigations of floodplain and lake settlements. J. Archaeol. Sci. 33, 14–33 (2006).Article 

Google Scholar 
Kittel, P. et al. A multi-proxy reconstruction from Lutomiersk-Koziówki, Central Poland, in the context of early modern hemp and flax processing. J. Archaeol. Sci. 50, 318–337 (2014).Article 

Google Scholar 
Kittel, P. et al. On the border between land and water: the environmental conditions of the Neolithic occupation from 4.3 until 1.6 ka BC at Serteya, Western Russia. Geoarchaeology 36, 173–202 (2021).Article 

Google Scholar 
Makohonienko, M. et al. Environmental changes during Mesolithic-Neolithic transition in Kuyavia Lakeland, Central Poland. Quat. Int. https://doi.org/10.1016/j.quaint.2021.11.020 (2021).Article 

Google Scholar 
Porinchu, D. F. & MacDonald, G. M. The use and application of freshwater midges (Chironomidae: Insecta: Diptera) in geographical research. Prog. Phys. Geogr. 27, 378–422 (2003).Article 

Google Scholar 
Brooks, S. J., Langdon, P. G. & Heiri, O. The Identification and Use of Palaearctic Chironomidae Larvae in Palaeoecology. QRA Technical guide no. 10 (Quaternary Research Association, 2007).Heiri, O., Birks, H. J. B., Brooks, S. J., Velle, G. & Willassen, E. Effects of within-lake variability of fossil assemblages on quantitative chironomid-inferred temperature reconstruction. Palaeogeogr. Palaeoclimatol. Palaeoecol. 199, 95–106 (2003).Article 

Google Scholar 
Kittel, P., Sikora, J. & Wroniecki, P. The morphology of the Luciąża River valley floor in the vicinity of the Rozprza medieval ring-fort in light of geophysical survey. Bull. Geogr. Phys. Geogr. Ser. 8, 95–106 (2015).Article 

Google Scholar 
Hingham, R. & Barker, P. Timber Castles (University of Exeter Press, 2002).
Google Scholar 
Marciniak-Kajzer, A. Archaeology on Medieval Knights’ Manor Houses in Poland (Wyd. Uniwersytetu Łódzkiego, Wyd. Uniwersytetu Jagiellońskiego, 2016).Book 

Google Scholar 
Moller Pillot, H. K. M. Chironomidae Larvae of the Netherlands and Adjacent Lowlands. Biology and Ecology of the Aquatic Orthocladiinae, Prodiamesinae, Diamesinae, Buchonomyiinae, Podonominae, Telmatogetoninae (KNNV Publishing, 2013).Book 

Google Scholar 
Luoto, T. P. An assessment of lentic ceratopogonids, ephemeropterans, trichopterans and oribatid mites as indicators of past environmental change in Finland. Ann. Zool. Fenn. 46, 259–270 (2009).Article 

Google Scholar 
Cierniewski, J. Spatial complexity of the Cybina river valley organic soils against the background of physiographic conditions. Soil Sci. Annu. 32(4), 3–51 (1981).CAS 

Google Scholar 
Rydelek, P. Origin and composition of mineral constituents of fen peats from Eastern Poland. J. Plant Nutr. 36(6), 911–928 (2013).Article 
CAS 

Google Scholar 
Wachecka-Kotkowska, L. Rozwój rzeźby obszaru między Piotrkowem Trybunalskim, Radomskiem a Przedborzem w czwartorzędzie (Wyd. Uniwersytetu Łódzkiego, 2015).Book 

Google Scholar 
Kittel, P. et al. Lacustrine, fluvial and slope deposits in the wetland shore area in Serteya, Western Russia. Acta Geogr. Lodz 110, 103–124 (2020).
Google Scholar 
Ciszewski, D. Pollution of Mała Panew River sediments by heavy metals: Part I. Effect of changes in river bed morphology. Pol. J. Environ. Stud. 13(6), 589–595 (2004).CAS 

Google Scholar 
Borówka, R. Late Vistulian and Holocene denudation magnitude in morainic plateaux: Case studies in the zone of maximum extent of the last ice sheet. Quat. Stud. Pol. 9, 5–31 (1990).
Google Scholar 
Prusinkiewicz, Z., Bednarek, R., Kośko, A. & Szmyt, M. Palaeopedological studies of the age and properties of illuvial bands at an archaeological site. Quat. Int. 51(52), 195–201 (1998).Article 

Google Scholar 
Kühtreiber, T. The medieval castle Lanzenkirchen in Lower Austria: reconstruction of economical and ecological development of an average-sized manor (12th–15th century). Archaeol. Pol. 37, 135–144 (1999).
Google Scholar 
Kočár, P., Čech, P., Kozáková, R. & Kočárová, R. Environment and economy of the early medieval settlement in Žatec. Interdiscip. Archaeol. 1, 45–60 (2010).
Google Scholar 
Brown, A. D. & Pluskowski, A. G. Detecting the environmental impact of the Baltic Crusades on a late medieval (13th-15th century) frontier landscape: Palynological analysis from Malbork Castle and hinterland, Northern Poland. J. Archaeol. Sci. 38, 1957–1966 (2011).Article 

Google Scholar 
Beneš, J. et al. Archaeobotany of the Old Prague Town defence system, Czech Republic: Archaeology, macro-remains, pollen, and diatoms. Veg. Hist. Archaeobot. 11(1/2), 107–119 (2002).Article 

Google Scholar 
Badura, M. & Latałowa, M. Szczątki makroskopowe roślin z obiektów archeologicznych Zespołu Przedbramia w Gdańsku. In Zespół Przedbramia ul. Długiej w Gdańsku. Studium archeologiczne (ed. Pudło, A.) 231–247 (Muzeum Historii Miasta Gdańska, 2016).
Google Scholar 
Dobrowolski, R. et al. Environmental conditions of settlement in the vicinity of the mediaeval capital of the Cherven Towns (Czermno site, Hrubieszów Basin, Eastern Poland). Quat. Int. 493, 258–273 (2018).Article 

Google Scholar 
Makohonienko, M. Środowisko przyrodnicze i gospodarka w otoczeniu średniowiecznego grodu w Łęczycy w świetle analizy palinologicznej. In Początki Łęczycy. Tom I—Archeologia środowiskowa średniowiecznej Łęczycy. Przyroda–Gospodarka–Społeczeństwo (eds Grygiel, R. & Jurek, T.) 95–190 (MAiE w Łodzi, 2014).
Google Scholar 
Koszałka, J. Źródła archeobotaniczne do rekonstrukcji uwarunkowań przyrodniczych oraz gospodarczych grodu w Łęczycy. In Początki Łęczycy. Tom I – Archeologia środowiskowa średniowiecznej Łęczycy. Przyroda–Gospodarka–Społeczeństwo (eds Grygiel, R. & Jurek, T.) 191–241 (MAiE w Łodzi, 2014).
Google Scholar 
Digerfeldt, G. Studies on past lake-level fluctuations. In Handbook of Holocene Palaeoecology and Palaeohydrology (ed. Berglund, B. E.) 127–143 (Wiley, 1986).
Google Scholar 
Magny, M. Palaeoclimatology and archaeology in the wetlands. In The Oxford Handbook of Wetland Archaeology (eds Menotti, F. & O’Sullivan, A.) 585–597 (Oxford University Press, 2013).
Google Scholar 
Płóciennik, M. et al. Summer temperature drives the lake ecosystem during the Late Weichselian and Holocene in Eastern Europe: A case study from East European Plain. CATENA 214, 106206 (2022).Article 

Google Scholar 
Święta-Musznicka, J., Badura, M., Pędziszewska, A. & Latałowa, M. Environmental changes and plant use during the 5th–14th centuries in medieval Gdańsk, northern Poland. Veget. Hist. Archaeobot. 30, 363–381 (2021).Article 

Google Scholar 
Rackham, J. & Sidell, J. London’s landscapes: The changing environment. In The Archaeology of Greater London. An Assessment of Archaeological Evidence for Human Presence in the Area Now Covered by Greater London (ed. Kendall, M.) 12–27 (Museum of London, 2000).
Google Scholar 
Ledger, P., Edwards, K. & Schofield, J. A multiple profile approach to the palynological reconstruction of Norse landscapes in Greenland’s Eastern Settlement. Quat. Res. 82(1), 22–37 (2014).Article 

Google Scholar 
Albert, B. & Innes, J. Multi-profile fine-resolution palynological and micro-charcoal analyses at Esklets, North York Moors, UK, with special reference to the Mesolithic-Neolithic transition. Veget. Hist. Archaeobot. 24, 357–375 (2015).Article 

Google Scholar 
Sikora, J., Kittel, P. & Wroniecki, P. From a point on the map to a shape in the landscape. Non-invasive verification of medieval ring-forts in Central Poland: Rozprza case study. Archaeol. Pol. 53, 510–514 (2015).
Google Scholar 
Sikora, J. et al. A palaeoenvironmental reconstruction of the rampart construction of the medieval ring-fort in Rozprza, Central Poland. Archaeol. Anthropol. Sci. 11(8), 4187–4219 (2019).Article 

Google Scholar 
Tolksdorf, J. F., Turner, F., Nelle, O., Peters, S. & Bruckner, H. Environmental development and local human impact in the Jeetzel valley (N Germany) since 10 ka BP as detected by geoarchaeological analyses in a coupled aeolian and lacustrine sediment archive at Soven. E&G Quat. Sci. J. 64, 95–110 (2015).Article 

Google Scholar 
Oonk S., Slomp C. P. & Huisman D. J. Geochemistry as an aid in archaeological prospection and site interpretation: Current issues and research directions. Archaeol. Prospect. 16, 35–51 (2009).Article 

Google Scholar 
Zieliński, T. & Pisarska-Jamroży, M. Which features of deposits should be included in a code and which not? Przegl. Geol. 60, 387–397 (2012).
Google Scholar 
Clift, P. D. et al. Grain-size variability within a mega-scale point-bar system, False River, Louisiana. Sedimentology 66, 408–434 (2019).Article 

Google Scholar 
Blott, S. J. & Pye, K. GRADISTAT: A grain size distribution and statistics package for the analysis of unconsolidated sediments. Earth Surf. Process. Landf. 26, 1237–1248 (2001).Article 

Google Scholar 
Rolland, N. & Larocque, I. The efficiency of kerosene flotation for extraction of chironomid head capsules from lake sediments samples. J. Paleolimnol. 37, 565–572 (2007).Article 

Google Scholar 
Schmid, P. E. A Key to the Chironomidae and Their Instars from Austrian Danube Region Streams and Rivers. Part I. Diamesinae Prodiamesinae and Orthocladiinae (Federal Institute for Water Quality of the Ministry of Agriculture and Forestry, 1993).
Google Scholar 
Andersen, T., Cranston, P. S. & Epler, J. H. Chironomidae of the Holarctic Region: Keys and Diagnoses. Part 1. Larvae. Insect Systematics and Evolution. Supplement 66 (Scandinavian Entomology, 2013).
Google Scholar 
Walker, I. R. Midges: Chironomidae and related Diptera. In Tracking Environmental Change Using Lake Sediments, Volume 4: Zoological Indicators (eds Smol, J. P. et al.) 43–66 (Kluwer Academic Press, 2001).Chapter 

Google Scholar 
Vallenduuk, H. J. & Moller Pillot, H. K. M. Chironomidae Larvae of the Netherlands and Adjacent Lowlands. General Ecology and Tanypodinae (KNNV Publishing, 2007).
Google Scholar 
Moller Pillot, H. K. M. Chironomidae Larvae Biology and Ecology of the Chironomini (KNNV Publishing, 2009).Book 

Google Scholar 
Juggins, S. C2 Version 1.5 User Guide. Software for Ecological and Palaeoecological Data Analysis and Visualisation (Newcastle University, 2007).
Google Scholar 
Schweingruber, F. H. Tree Rings. Basics and Applications of Dendrochronology (Kluwer Academic Publishers, 1988).
Google Scholar 
Skripkin, V. V. & Kovaliukh, N. N. Recent developments in the procedures used at the SSCER Laboratory for the routine preparation of lithium carbide. Radiocarbon 40(1), 211–214 (1998).Article 
CAS 

Google Scholar 
Krąpiec, M., Rakowski, A. Z., Huels, M., Wiktorowski, D. & Hamann, C. A new graphitization system for radiocarbon dating with AMS on the dendrochronological laboratory at AGH-UST Kraków. Radiocarbon 60(4), 1091–1100 (2018).Article 

Google Scholar 
Zoppi, U., Crye, J., Song, Q. & Arjomand, A. Performance evaluation of the new AMS system at Accium BioSciences. Radiocarbon 49, 173–182 (2007).Article 
CAS 

Google Scholar 
Reimer, P. et al. The IntCal20 Northern Hemisphere radiocarbon age calibration curve (0–55 cal kBP). Radiocarbon 62(4), 725–757 (2020).Article 
CAS 

Google Scholar 
Bronk Ramsey, C. OxCal Version 4.4.2. Available at: https://c14.arch.ox.ac.uk (2020).Bronk Ramsey, C. Bayesian analysis of radiocarbon dates. Radiocarbon 51(1), 337–360 (2009).Article 

Google Scholar 
Bronk Ramsey, C. Deposition models for chronological records. Quat. Sci. Rev. 27(1–2), 42–60 (2008).Article 

Google Scholar 
Kohonen, T. Self-organized formation of topologically correct feature maps. Biol. Cybern. 43, 59–69 (1982).Article 
MathSciNet 
MATH 

Google Scholar 
Kohonen, T. Self-Organizing Maps (Springer, 2001).Book 
MATH 

Google Scholar 
Park, Y.-S. et al. Application of a self-organizing map to select representative species in multivariate analysis: A case study determining diatom distribution patterns across France. Ecol. Inform. 1, 247–257 (2006).Article 

Google Scholar 
Zhang, Q. et al. Self-organizing feature map classification and ordination of Larix principis-rupprechtii forest in Pangquangou Nature Reserve. Acta Ecol. Sin. 31, 2990–2998 (2011).
Google Scholar 
Ney, J. J. Practical use of biological statistics. In Inland Fisheries Management in North America (eds Kohler, C. C. et al.) 137–158 (American Fisheries Society, 1993).
Google Scholar 
Płóciennik, M. et al. Fen ecosystem responses to water-level fluctuations during the early and middle Holocene in central Europe: A case study from Wilczków, Poland. Boreas 44(4), 721–740 (2015).Article 

Google Scholar 
Brosse, S., Giraudel, J. L. & Lek, S. Utilisation of non-supervised neural networks and principal component analysis to study fish assemblages. Ecol. Model. 146(1), 159–166 (2001).Article 

Google Scholar 
Lek, S., Scardi, M., Verdonschot, P. F. M., Descy, J. P. & Park, Y. S. Modelling Community Structure in Freshwater Ecosystems (Springer, 2005).Book 

Google Scholar 
Quinn, G. P. & Keough, M. Experimental Design and Data Analysis for Biologists (University of Cambridge, 2002).Book 

Google Scholar 
Płóciennik, M., Kruk, A., Michczyńska, D. J. & Birks, H. J. B. Kohonen artificial neural networks and the IndVal index as supplementary tools for the quantitative analysis of palaeoecological data. Geochronometria 42, 189–201 (2015).Article 

Google Scholar 
Vesanto, J. & Alhoniemi, E. Clustering of the self-organizing map. IEEE Trans. Neural Netw. 11, 586–600 (2000).Article 
CAS 
PubMed 

Google Scholar 
Ward, J. H. Hierarchical grouping to optimize an objective function. J. Am. Stat. Assoc. 58, 236–244 (1963).Article 
MathSciNet 

Google Scholar 
Alhoniemi, E., Hollmén, J., Simula, O. & Vesanto, J. Process monitoring and modeling using the self-organizing map. Integr. Comput. Aided Eng. 6(1), 3–14 (1999).Article 

Google Scholar 
Dufrêne, M. & Legendre, P. Species assemblages and indicator species: The need for a flexible asymmetrical approach. Ecol. Monogr. 67, 345–366 (1997).
Google Scholar 
McCune, B. & Mefford, M. S. PcOrd Multivariate Analysis of Ecological Data. Version 6.06 (MjM Software, 2011).
Google Scholar 
Hadfield, J. D., Krasnov, B. R., Poulin, R. & Nakagawa, S. A tale of two phylogenies: Comparative analyses of ecological interactions. Am. Nat. 183(2), 174–187 (2014).Article 
PubMed 

Google Scholar 
R Core Team. R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, 2021).
Google Scholar 
Brooks, M. E. et al. glmmTMB balances speed and flexibility among packages for zero-inflated generalized linear mixed modeling. R Journal 9(2), 378–400 (2017).Article 

Google Scholar 
Hartig, F. DHARMa: Residual Diagnostics for Hierarchical (Multi-level/Mixed) Regression Models. R Package Version 0.4.5. https://CRAN.R-project.org/package=DHARMa (2022).Bartoń, K. MuMIn: Multi-model Inference. R Package Version 1.43.17. https://CRAN.R-project.org/package=MuMIn (2020).de Rosario-Martinez, H. phia: Post-Hoc Interaction Analysis. R Package Version 0.2-1. https://CRAN.R-project.org/package=phia (2015). More