Appendix S1 SUPPORTING INFORMATION Europe’s freshwater biodiversity under climate change: distribution shifts and conservation needs Danijela Markovic, Savrina Carrizo, Jörg Freyhof, Nuria Cid, Lengyel Szabolcs, Mathias Scholz, Hans Kasperidus, William Darwall a) b) Figure S1: a) A comparison of the different resolutions of the HydroBasins dataset (level 4, level 6 and level 8) for the Elbe River Basin (green); b) distribution of species occurrence frequencies (defined as the total number of positive species occurrences (presence in a catchment=1, absence=0) across the studied HydroBasin level 8 catchments (18 783 European catchments). Remarks: a) The average catchment area at the HydroBasins level 8 resolution is 536.3 km2. Due to the small size of the catchments involved (on average about 25 km x 20 km), the parameters such as the mean catchment temperature and the altitudinal range hold too little variability to be meaningful for modelling distributions of freshwater species. b) Notice that the proportion of species occurring in 30-40, 40-50, 50-60 and 60-70 catchments is between 1% and 2% of the total species data sample. Due to a small number of species having between 20 and 50 and between 50 and 70 occurrences, using slightly higher or lower values than 50 as a threshold for the SDM application would not change our results as only 1-2% of species would be affected. 1 Figure S2: Parameter correlation matrix Remarks: Given that the analyses in our current manuscript involved calculating spatial statistics for about 19 000 catchments, we restricted the extraction of the catchment related worldclim predictor dataset to Altitude, AnnTMean (mean annual temperature), AnnTMax (maximum temperature of warmest month) and AnnPMean (mean annual precipitation), plus the mean winter temperature (WinterT), mean summer temperature (SummerT), temperature seasonality (TSeason), precipitation of the driest quarter (DryQP), temperature of the driest quarter (DryQT) as well as latitude and GDP (the latter representing anthropogenic pressure). Since our analysis units were catchments and not sites, we realised that the use of mean temperatures (AnnTMean, WinterT, SummerT, DryQT) cannot reflect the temperatures that species may experience when dispersing though stream networks of the catchments. Therefore, we focussed here on AnnTMax. As illustrated in the predictor correlation matrix, the number of statistically applicable factors (not violating the multicollinearity condition, i.e. correlation <0.7) was therefore limited to exactly those we used (with GDP and Lat excluded to avoid overfitting). 2 Figure S3: Illustration of the monthly thermal niche approach. Min/max are the min and max monthly temperatures across a current distribution range of a hypothetical species; the green and grey squares denote the predicted mean monthly temperatures for the catchments A and B, respectively according to a single climate model. Following the monthly thermal niche approach the catchment A would be considered “suitable” (for that particular climate model) while the catchment B would be considered “not-suitable”. If the same conclusion (“suitable”) is obtained for all climate models, then a catchment is considered “suitable, otherwise, “notsuitable”. 3 SDMs monthly thermal niche approach a) Common species, no dispersal scenario SDMs monthly thermal niche approach b) Common species, free-dispersal scenario SDM- based monthly thermal niche approach c) Rare species, no-dispersal scenario monthly thermal niche approach SDM- based d) Rare species, free-dispersal scenario Figure S4. A comparison of the species richness of 2050s distribution patterns for common (a-b) and rare (c-d) freshwater species across European catchments for the no-dispersal scenario and the free-dispersal scenario using alternative approaches: SDMs vs. monthly thermal niche approach (a-b) and SDM-parameters based vs. monthly thermal niche approach (c-d). Within the SDM- parameters based approach (c-d, left panel), a proxy for the current species’ environmental niche was estimated by calculating the minimum and the maximum across current species ranges of the parameters used in SDM-based modelling of common species distributions (maximum air temperature of the warmest month- AnnTMax, seasonal temperature variability-TSeason, the mean annual precipitation-AnnPMean and altitude). Within the SDM- parameters based approach a catchment is considered “suitable” for a particular species in the future only if, for all three climate models considered, the future predicted values of AnnTMax, TSeason and AnnPMean and the average catchment altitude are between the minimum and the maximum detected across current species ranges, otherwise it is classified as “not suitable”. Remarks: Compared to the SDMs, the monthly thermal niche approach provides conservative predictions of common freshwater species richness in southern Europe, and vice versa in central and northern Europe (a-b, right panel). Only for the no-dispersal scenario (a), there is a good agreement between the two approaches. The latter suggests that the no-dispersal scenario predictions of rare species distributions (see main text) is most likely comparable to the result that could be obtained by the SDMs if there would be enough positive species occurrences to run the models and are more reliable than the free-dispersal scenario predictions. Furthermore, compared to the monthly thermal niche approach (c-d), the SDM-parameters based approach is generally more conservative. Given the similarity of the patters obtained using SDMs and monthly niche approach for the nodispersal scenario (a), we conclude appropriateness of the latter approach to analyse rare species distributions. 4 Figure S5. Current and SDM- based future patterns of common freshwater species across European catchments: (a-c) fish, (d-f) molluscs, (g-i) plants, (j-l) odonates and (m-o) amphibians. The first figure in each row shows the current species richness, the second shows the projection for the 2050s climate assuming no dispersal, and the third shows the projection for the 2050s climate assuming species dispersal to suitable catchments, constrained by the hydrological catchment connectivity for fish, crayfish and molluscs. 5 Figure S6: Distribution of rare freshwater species across major European river basins: the bar plot shows the current proportion of freshwater species that occur in each basin, while the filled and empty squares indicate the proportion of species predicted to occur under 2050s climate based on the A1b emission scenario for the nodispersal and the free-dispersal scenario, respectively. Abbreviations for major European river basins are as follows: Danu-Danube, Daug-Daugava, Dnie-Dnieper, Dnis-Dnister Nistru, Don-Don, Duer-Duero, Ebro-Ebro, Elbe-Elbe, Evro-Evros/Maritsa, Garo-Garonne, Guaq-Guadalquivir, Guad-Guadiana, Loir-Loire, Meze-Mezen, Narv-Narva, Nemu-Nemunas, Neva-Neva, Nort-Northern Dvina, Oder-Oder, Pech-Pechora, Po-Po, Rhin-Rhine, Rhon-Rhone, Sein-Seine, Sout-Southern Bug, Tajo-Tajo, Tere-Terek, Ural-Ural, Volg-Volga, Wisl-Wisla. . 6 Figure S7: Distribution of rare freshwater species across European freshwater ecoregions: the bar plot shows the current proportion of freshwater species that occur in each ecoregion, while the filled and empty squares indicate the proportion of species predicted to occur under 2050s climate based on the A1b emission scenario for the nodispersal and the free-dispersal scenario, respectively. Abbreviations for freshwater ecoregions are as follows: AD-Aegean Drainages, BSD-Barents Sea Drainages, CC-Cantabric Coast - Languedoc, CM-Caspian Marine, CP-Crimea Peninsula, CWE-Central & Western Europe, DAL-Dalmatia, DLD-Dniester - Lower Danube, DONDon, DSB-Dnieper - South Bug, DU-Upper Danube, EI-Eastern Iberia, GVD-Gulf of Venice Drainages, ICIceland - Jan Mayen, ID-Ionian Drainages, IPI-Italian Peninsula & Islands, IT-Irgyz -Turgai, KSCD-Kura South Caspian Drainages, KUB-Kuban, LOL-Lake Onega - Lake Ladoga, NBD-Northern Baltic Drainages, NBI-Northern British Isles, NSD-Norwegian Sea Drainages, OB-Ob, SAD-Southeastern Adriatic Drainages, SBL-Southern Baltic Lowlands, SI-Southern Iberia, THR-Thrace, VAR-Vardar, VD-Volga Delta - Northern Caspian Drainages, VU-Volga - Ural, WA-Western Anatolia, WCD-Western Caspian Drainages, WI-Western Iberia, WT-Western Transcaucasia. 7 Figure S8: Distribution of common freshwater species across European countries (countries having area>2000km2 are considered): the bar plot shows the current proportion of common freshwater species that occur in each individual country, while the filled and empty squares indicate the proportion of species predicted to occur under 2050s climate based on the A1b emission scenario for the no-dispersal and the free-dispersal scenario, respectively. Country abbreviations are as follows: AL-Albania, AU-Austria, BE-Belgium, BK-Bosnia and Herzegovina, BO-Belarus, BU-Bulgaria, DA-Denmark, EI-Ireland, EN-Estonia, EZ-Czech Republic, FIFinland, FR-France, GM-Germany, GR-Greece, HR-Croatia, HU-Hungary, IC-Iceland, IT-Italy, LG-Latvia, LHLithuania, LO-Slovakia, LU-Luxembourg, MD-Moldova, MK-The Former Yugoslav Republic of Macedonia, MW-Montenegro, NL-Netherlands, NO-Norway, PL-Poland, PO-Portugal, RO-Romania, RS-Russia, SISlovenia, SP-Spain, SR-Serbia, SV-Svalbard, SW-Sweden, SZ-Switzerland, UK-United Kingdom, UP-Ukraine. 8 Figure S9: Distribution of rare freshwater species across European countries (countries having area>2000km2 are considered): the bar plot shows the current proportion of freshwater species that occur in each individual country, while the filled and empty squares indicate the proportion of species predicted to occur under 2050s climate based on the A1b emission scenario for the no-dispersal and the free-dispersal scenario, respectively. Country abbreviations are as follows: AL-Albania, AU-Austria, BE-Belgium, BK-Bosnia and Herzegovina, BOBelarus, BU-Bulgaria, DA-Denmark, EI-Ireland, EN-Estonia, EZ-Czech Republic, FI-Finland, FR-France, GMGermany, GR-Greece, HR-Croatia, HU-Hungary, IC-Iceland, IT-Italy, LG-Latvia, LH-Lithuania, LO-Slovakia, LU-Luxembourg, MD-Moldova, MK-The Former Yugoslav Republic of Macedonia, MW-Montenegro, NLNetherlands, NO-Norway, PL-Poland, PO-Portugal, RO-Romania, RS-Russia, SI-Slovenia, SP-Spain, SRSerbia, SV-Svalbard, SW-Sweden, SZ-Switzerland, UK-United Kingdom, UP-Ukraine. 9 Figure S10: Catchments with non-analogue climates by 2050s (red). A catchment is considered to have a nonanalogue climate by 2050s, if at least one of the used global circulation models (ECHAM5, IPSL and HadCM3) predicted higher values for any of the climatic variables used to calibrate species distribution models (maximum temperature of the warmest month-AnnTMax; seasonal variability in temperature-TSeason; mean annual precipitation- AnnPMean) than the overall climate extremes for the 20 th century climate across the study area (AnnTMaxbaseline,max= 43.7 °C; TSeasonbaseline,max=1.4 °C; AnnPMeanbaseline,max=2306 mm). a) b) Figure S11: (a) The European protected area network (green, Natura 2000 and WDPAs with IUCN categories IIV) and (b) the areas that are currently suitable and predicted to be suitable by the 2050s (blue) for the “gap” species retaining parts of their current range in the future (see Table S4-S5 for species list) and the urgent priority catchments for these species (red). We considered a catchment to be an “urgent priority catchment” (UPC) for conservation if it is currently unprotected, but suitable for a “gap species” under current and future (2050s) climate, and, if protected, would thus ensure that at least 100km2 of current species range area would be protected. Remarks: Given the greater density of protected areas in western Europe, it is not surprising that the urgent priority catchments are mainly located in the eastern parts of Europe (mainly Russia) and Balkan- countries (Croatia and Montenegro. Note, the UPCs only indicate the catchments that would ensure that a minimum of 100km2 of the range area suitable under the current- and 2050s climate under the no-dispersal scenario would be protected for the identified “gap” species. As the UPCs do not consider species range shifts, they are a suitable conservation strategy for only a minority of the gap-species (34 species, see Table S4 and S5). An analysis of the co-occurrence of these 34 species within the areas identified as currently suitable and suitable in the future, has revealed that protection measures across only 12 catchments (UPCs area =7 783km2, red area in b) would assure species removal from the “gap list”. 10 a) b) Figure S12: Boxplots of the Area Under the receiver operating Curve (AUC) a) and of the True Skill Statistic (TSS) b) per taxa group studied (the least represented taxa, crayfish and turtles, have been excluded from the analysis). An AUC of 0.5 indicates that a model has no discriminatory power, while an AUC of 1 indicates that “suitable” and “not suitable” catchments are perfectly discriminated. A TSS of 0 indicates that a model has no discriminatory power, while a TSS of 1 indicates that “suitable” and “not suitable” catchments are perfectly discriminated. The bottom and top of the box denote the first and third quartiles, the line inside the box denote the median, while whiskers denote the extreme values. 11 Table S1: The number of European freshwater species predicted to lose more than 90% of their current range Plants Fish Molluscs Odonates Amphibians common species 5 23 19 3 0 rare species 15 178 381 24 5 totals [%] 6% 39% 61% 20% 9% Table S2: Rare freshwater species predicted to lose suitability across the entire current range area by 2050s, paired with no suitable catchments in Europe by 2050s. CR=Critically Endangered, EN=Endangered, VU=Vulnerable, NT=Near Threatened, LC=Least Concern, DD=Data Deficient, NA=Not Assessed. Scientific name Alburnus macedonicus Alosa killarnensis Alosa macedonica Alosa sp nov Skadar Alosa vistonica Benthophilus mahmudbejovi Coregonus atterensis Coregonus baerii Coregonus bavaricus Coregonus danneri Coregonus fontanae Coregonus hoferi Coregonus ladogae Coregonus lucinensis Coregonus lutokka Coregonus pollan Economidichthys trichonis Gymnocephalus ambriaelacus Hypomesus olidus Knipowitschia cameliae Ponticola eurycephalus Rhynchocypris czekanowskii Rutilus meidingeri Salaria economidisi Salvelinus evasus Salvelinus gracillimus Salvelinus murta Salvelinus obtusus Salvelinus struanensis Salvelinus thingvallensis Squalius albus Coenagrion ecornutum Elatine gussonei Zannichellia melitensis Isoetes boryana Callitriche transvolgensis Apium bermejoi Rorippa valdes bermejoi Taxa group fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish fish odonates plants plants plants plants plants plants Red List Status (European) CR CR VU VU CR LC VU DD CR VU LC CR LC VU LC EN EN CR NA CR LC NA EN CR VU VU LC CR VU LC LC NA LC LC EN NA CR CR 12 Potamogeton epihydrus Belgrandia alcoaensis Belgrandia conoidea Belgrandia latina Bithynia hambergerae Bithynia skadarskii Bithynia zeta Bracenica spiridoni Bythinella carinulata Bythinella jourdei Bythinella micherdzinskii Bythinella viridis Bythinella zyvionteki Bythiospeum acicula Bythiospeum haessleini Gyraulus meierbrooki Heleobia dobrogica Horatia lucidulus Islamia lagari Marstoniopsis armoricana Mercuria sarahae Mercuria vindilica Paladilhia pontmartiniana Plagigeyeria deformata Pseudamnicola bacescui Pseudamnicola leontina Pseudobithynia ambrakis Pyrgula annulata Radix skutaris Radomaniola elongata Radomaniola lacustris Salenthydrobia ferrerii Spiralix collieri Stagnicola montenegrinus Theodoxus baeticus Turricaspia chersonica Turricaspia ismailensis Unio gibbus plants molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs molluscs NT CR EN VU DD EN EN EN EN VU VU EN EN VU VU EN CR CR VU CR CR EN DD EN VU DD VU LC EN CR CR EN DD NT CR NA VU CR 13 Table S3: Current species richness and the 2050s perspective for common freshwater species across European countries. Projections for 2050s are given in terms of proportion (%) of species that may occur in each individual country for the no-dispersal and the free-dispersal scenario. Plants Albania Austria Belarus Belgium Bosnia and Herz. Bulgaria Croatia Czech Republic Denmark Estonia Finland France Germany Greece Hungary Iceland Ireland Italy Latvia Lithuania Luxembourg Moldova Montenegro Netherlands Norway Poland Portugal Romania Russia Serbia Slovakia Slovenia Spain Svalbard Sweden Switzerland FYR Macedonia Ukraine United Kingdom c nd 187 229 216 190 181 202 220 209 176 173 178 254 224 198 221 64 167 244 182 188 184 191 183 188 177 220 194 211 239 206 215 217 232 11 188 214 192 218 188 -43 -13 -13 -10 -32 -38 -33 -12 -5 -14 -3 -13 -9 -45 -37 -5 -7 -23 -12 -11 -11 -39 -37 -12 -6 -11 -18 -19 -3 -44 -17 -34 -22 -27 -2 -14 -52 -12 -5 Fish fd c -35 48 -6 70 -5 55 5 50 -26 73 -30 90 -26 86 -4 68 7 44 14 42 30 49 -6 91 -1 75 -30 54 -29 62 27 7 10 26 -18 90 12 47 3 49 2 36 -31 80 -35 64 5 55 18 47 -4 73 -15 38 -15 88 -1 121 -35 79 -8 68 -26 73 -12 67 104 1 19 52 -4 60 -40 73 -10 102 10 39 Molluscs nd fd -33 -10 -13 -14 -22 -27 -26 -24 -11 -10 -8 -11 -11 -17 -29 0 -8 -23 -6 -14 -25 -18 -25 -20 -6 -10 -8 -10 -3 -29 -21 -26 -27 0 -8 -15 -45 -11 -5 -23 6 2 6 -23 -26 -29 11 -7 12 34 -6 6 -25 -22 0 9 -11 9 -8 -7 -9 -21 -10 2 9 -12 -8 2 -28 1 -16 -22 0 -4 -7 -34 -2 3 c nd Odonates Amphibians all taxa fd c nd fd c nd fd c nd fd 68 -26 -29 106 -11 -3 76 -9 -4 83 -8 -2 74 -16 -27 75 -19 -22 101 -28 -25 84 -7 -1 81 -7 4 69 -7 -4 68 -4 17 111 -12 -9 98 -2 4 74 -36 -33 90 -38 -24 9 0 0 55 -5 4 124 -19 -12 72 0 4 75 -4 -1 72 -10 -6 76 -24 -14 68 -10 -23 82 -9 -5 70 -3 1 86 -3 8 58 -5 -5 82 -23 -25 86 -1 1 84 -33 -28 84 -18 -6 101 -30 -26 95 -19 -16 1 -100 0 71 -1 4 93 -12 -4 73 -38 -22 86 -12 -1 77 -1 -1 65 81 70 69 69 70 76 77 62 59 55 96 78 72 73 1 32 97 64 67 67 63 63 68 47 74 67 70 85 69 76 76 86 0 60 78 69 75 59 -9 -12 -9 -13 -14 -13 -14 -14 -6 -8 -4 -8 -4 -19 -23 0 -13 -10 -8 -4 -10 -17 -8 -19 -2 -5 -9 -14 -1 -14 -17 -22 -16 0 -3 -12 -17 -11 -5 -2 4 1 10 -5 -7 -13 1 49 27 33 1 17 -9 -7 0 122 -3 19 12 13 -2 -5 3 27 9 -6 -10 0 -1 -6 -10 -6 200 22 6 -4 -1 19 14 20 13 20 16 18 21 19 13 12 5 29 21 16 17 0 3 27 13 13 17 13 13 19 6 18 18 18 16 18 18 18 25 0 12 22 13 19 19 -7 -15 0 -10 -31 -33 -33 -5 -8 0 0 -10 -5 -31 -35 0 -33 -30 0 0 0 -38 0 -11 0 0 -6 -17 0 -17 -17 -22 -16 0 -8 -18 -8 -5 -11 -17 19 31 32 -12 -24 -21 32 122 50 70 5 35 -18 -9 100 220 -16 80 29 29 -7 4 15 44 39 -11 -9 0 -6 21 -17 -3 0 47 13 -24 26 28 388 513 434 421 420 460 512 464 381 358 359 592 506 419 468 81 286 592 382 396 385 427 398 418 351 478 379 474 553 462 466 492 513 13 386 477 427 506 388 -31 -12 -11 -11 -24 -28 -28 -13 -6 -11 -4 -11 -7 -34 -34 -4 -7 -20 -8 -8 -12 -29 -24 -13 -5 -8 -12 -17 -2 -34 -18 -29 -20 -31 -3 -13 -41 -11 -4 -25 0 -2 6 -21 -24 -24 1 13 13 30 -5 6 -25 -22 23 23 -13 13 3 2 -18 -23 1 16 4 -11 -15 0 -25 -5 -21 -12 103 15 -1 -28 -4 10 The “c”, “nd, and “fd” denote current species richness, proportion (%) of species projected for 2050s following the no-dispersal scenario and, proportion of species projected for 2050s following the free-dispersal scenario, respectively. The columns “all taxa” summarize the results across all species groups (plants, fish, molluscs, odonates, amphibians, crayfish and turtles). 14 Table S4: List of common freshwater “gap” species (species in urgent priority catchments are marked with an asterisk). We considered a catchment to be an “urgent priority catchment” (UPC) for conservation if it is currently unprotected, but suitable for a “gap species” under current and future (2050s) climate, and, if protected, would thus ensure that at least 100km2 of current species range area would be protected (see Fig. S7). Scientific name Acipenser sturio Barbus kubanicus * Barbus waleckii Benthophilus durrelli * Chondrostoma kubanicum Clupeonella caspia Red List Status (European) Fish NA Fish LC Fish LC Fish LC Fish LC * Fish LC * Fish NA Fish NA Coregonus muksun Coregonus nasus * Taxa * Coregonus pallasii Fish LC * Fish LC * Fish LC Gobio occitaniae Fish LC Gobio sarmaticus Fish LC * Fish LC Fish LC Fish LC Fish LC Fish LC Fish LC Belgrandia minuscula Molluscs DD Bythinella ferussina Molluscs LC Bythinella simoniana Molluscs LC Fissuria boui Molluscs NT Islamia piristoma Molluscs LC Moitessieria locardi Molluscs LC Moitessieria simoniana Molluscs LC Theodoxus prevostianus Molluscs EN Turricaspia lindholmiana Molluscs NA Odonates LC Odonates NA Odonates DD Plants NA Plants NA Gobio brevicirris Gobio kubanicus Gobio volgensis Lethenteron reissneri Leuciscus danilewskii * Leuciscus oxyrrhis Luciobarbus albanicus Sabanejewia kubanica Aeshna crenata * * Somatochlora graeseri * Somatochlora sahlbergi Trapa alatyrica * Zannichellia clausii * * 15 Table S5: List of rare freshwater “gap” species in urgent priority catchments. We considered a catchment to be an “urgent priority catchment” (UPC) for conservation if it is currently unprotected, but suitable for a “gap species” under current and future (2050s) climate, and, if protected, would thus ensure that at least 100km 2 of current species range area would be protected. Salmo obtusirostris Red List Status (European) Fish Alburnoides kubanicus Fish LC Squalius tenellus Fish EN Chondrostoma knerii Fish VU Squalius svallize Fish VU Phoxinellus alepidotus Fish EN Chondrostoma phoxinus Fish EN Scardinius dergle Fish NT Cobitis dalmatina Fish VU Cobitis narentana Fish VU Phoxinellus pseudalepidotus Fish VU Salmo ezenami Fish CR Bithynia mostarensis Molluscs DD Theodoxus subterrelictus Molluscs EN Saxurinator sketi Molluscs EN Paladilhiopsis solida Molluscs DD Plagigeyeria mostarensis Molluscs DD Lanzaia kotlusae Molluscs VU Scientific name Red List Status (European) EN 16