Supplementary Information. Integrated trophic position decreases in more diverse

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1 Supplementary Information Integrated trophic position decreases in more diverse communities of stream food webs Naoto F. Ishikawa 1,*,, Yoshito Chikaraishi 1,2, Naohiko Ohkouchi 1, Aya R. Murakami 3, Ichiro Tayasu 4, Hiroyuki Togashi 5, Jun-ichi Okano 3, Yoichiro Sakai 6, Tomoya Iwata 7, Michio Kondoh 8, Noboru Okuda 4 1 Japan Agency for Marine-Earth Science and Technology, 2-15 Natsushima-cho, Yokosuka, Kanagawa , Japan 2 Institute of Low Temperature Science, Hokkaido University, Kita 19, Nishi 8, Kita-ku, Sapporo, Japan 3 Center for Ecological Research, Kyoto University, Hirano , Otsu, Shiga , Japan 4 Research Institute for Humanity and Nature, Motoyama, Kamigamo, Kita-ku, Kyoto, Japan 5 Tohoku National Fisheries Research Institute, Japan Fisheries Research and Education Agency, , Shinhama-cho, Shiogama, Miyagi , Japan 6 Lake Biwa Environmental Research Institute, 5-34 Yanagasaki, Otsu, Shiga , Japan 7 University of Yamanashi, Takeda, Kofu, Yamanashi , Japan 8 Department of Environmental Solution Technology, Ryukoku University, 1-5 Yokoya, Seta Oe-cho, Otsu, Shiga , Japan *Correspondence to: naoto.f.ishikawa@gmail.com Current address: ETH Zürich, Sonneggstrasse Zürich, Switzerland 1

2 Fig. S1. δ 15 N Glu (shaded square) and δ 15 N Phe (open square) values, and the itp values (circle) calculated for macroinvertebrate communities collected from 15 sites on the Yasu River. Open circles indicate stream orders 1 or 2, and shaded circles indicate stream orders 3 or 4. δ 15 N of glutamic acid ( ) itp (stream orders: 3 or 4) δ 15 N of phenylalanine ( ) itp (stream orders: 1 or 2) 3 2 itp δ 15 N ( )

3 Fig. S2. Relationships of (a) species density (m 2 ) or (b) itp to the number of individuals (m 2 ). The enlarged right panels exclude one sample with an extremely large number of individuals (i.e. site 10). Relationships between the itp and (c) -based Shannon Wiener index (H') or (d) individual-based Pielou evenness index (J'). Open circles indicate stream orders 1 or 2, and shaded circles indicate stream orders 3 or 4. (a) (b) Species density (m 2 ) itp Number of individuals (m 2 ) (c) Number of individuals (m 2 ) (d) itp 2.6 itp Diversity index H' (-based) Evenness index J' (individual-based) 3

4 Fig. S3. Relationships between integrated trophic position (itp) for macroinvertebrates and species density (m 2 ) and the individual-based Shannon Wiener diversity index (H') for the first replication (a and b, respectively) and those for the second replication (c and d, respectively). All correlations were significant (R 2 = , p < 0.01). Solid, dotted, and dashed lines represent the regression equation, 95% confidence interval, and 95% prediction interval, respectively. (a) (b) itp (c) Species density (m 2 ) Diversity index H' (individual-based) (d) itp Species density (m 2 ) Diversity index H' (individual-based) 4

5 Table S1. omic information. Individual (mg) was estimated by weighing several individuals from each taxon. Individual Individual Name of order Name of family Name of species # SD 1 Tricladida Dugesiidae Dugesia japonica Gordioida Chordodidae Chordodes sp Architaenioglossa Viviparidae Sinotaia quadrata histrica Discopoda Pleuroceridae Semisulcospira libertina Semisulcospira reiniana Basommatophora Lymnaeidae Fossaria ollula Physidae Physa acuta Veneroida Corbiculidae Corbicula sp Tubificida Naididae Branchiura sowerbyi Nais sp Naididae gen. sp Naididae gen. spp Lumbricida Megascolecidae Megascolecidae gen. sp Arhynchobdellida Erpobdellidae Erpobdella octoculata Erpobdella testacea Amphipoda Crangonyctidae Crangonyx floridanus Isopoda Asellidae Asellus hilgendorfi hilgendorfi Decapoda Atyidae Neocaridina denticulata Cambaridae Procambarus clarkii Potamidae Geothelphusa dehaani

6 Table S1. Continued. Individual Individual Name of order Name of family Name of species # SD 21 Ephemeroptera Baetidae Acentrella gnom Alainites yoshinensis Baetiella japonica Baetis sahoensis Baetis taiwanensis Baetis thermicus Baetis sp. J Baetis sp Labiobaetis atrebatinus orientalis Tenuibaetis flexifemora Heptageniidae Ecdyonurus yoshidae Epeorus curvatulus Epeorus sp Isonychiidae Isonychia japonica Leptophlebiidae Choroterpes altioculus Paraleptophlebia sp Ephemeridae Ephemera japonica Ephemera orientalis Ephemera strigata Potamanthidae Potamanthus formosus

7 Table S1. Continued. Individual Individual Name of order Name of family Name of species # SD 41 Ephemerellidae Cincticostella sp Ephemerella setigera Torleya japonica Uracanthella punctisetae Odonata Calopterygidae Calopteryx atrata Gomphidae Davidius sp Nihonogomphus viridis Onychogomphus viridicostus Cordulegasteridae Anotogaster sieboldii N/A N/A 50 Plecoptera Nemouridae Amphinemura sp Nemoura sp Protonemura sp Chloroperlidae Chloroperlidae gen. sp Perlidae Kamimuria sp Neoperla sp Oyamia sp N/A 57 Paragnetina sp Togoperla sp Perlinae gen. sp Hemiptera Gerridae Metrocoris histrio

8 Table S1. Continued. Individual Individual Name of order Name of family Name of species # SD 61 Aphelochiridae Aphelocheirus vittatus Megaloptera Corydalidae Protohermes grandis Neuroptera Nevrorthidae Nevrorthidae gen. sp Ttrichoptera Hydropsychidae Cheumatopsyche brevilineata Cheumatopsyche infascia Cheumatopsyche sp Diplectrona sp Hydropsyche albicephala Hydropsyche ancorapunctata Hydropsyche orientalis Philopotamidae Dolophilodes sp Stenopsychidae Stenopsyche marmorata Stenopsyche sauteri Stenopsyche sp. N/A N/A 75 Glossosomatidae Agapetus sp Glossosoma sp Glossosomatidae gen. spp Hydrobiosidae Apsilochorema sutshanum Rhyacophilidae Rhyacophila kawamurae Rhyacophila lezeyi

9 Table S1. Continued. Individual Individual Name of order Name of family Name of species # SD 81 Rhyacophila shikotsuensis Rhyacophila sp Goeridae Goera japonica Goera sp Lepidostomatidae Lepidostoma sp Leptoceridae Trichosetodes japonicus Sericostomatidae Gumaga orientalis Lepidoptera Crambidae Potamomusa midas Diptera Tipulidae Antocha sp Dicranota sp Hexatoma sp Tipula sp Chironomidae Brillia sp Cardiocladius sp Chironomus sp Cladotanytarsus sp Conchapelopia sp Cryptochironomus sp Cryptotendipes sp Dicrotendipes sp

10 Table S1. Continued. Individual Individual Name of order Name of family Name of species # SD 101 Eukiefferiella sp Microtendipes sp Orthocladius sp Orthocladius spp Chironomidae gen. sp Chironomidae gen. spp Parametriocnemus sp Polypedilum sp Potthastia longimana Pseudorthocladius sp Rheopelopia joganflava Rheotanytarsus sp. 0.1 N/A 113 Stictochironomus sp Tanytarsus sp Tvetenia sp Simuliidae Simulium sp Athericidae Atrichops morimotoi Stratiomyidae Stratiomyidae gen. sp Dolichopodidae Dolichopodidae gen. sp Coleoptera Hydrophilidae Laccobius oscillans

11 Table S1. Continued. Individual Individual Name of order Name of family Name of species # SD 121 Elmidae Ordobrevia maculata Zaitzevia nitida Zaitzeviaria brevis Elminae sp Elminae spp Psephenidae Ectopria opaca opaca Eubrianax granicollis Mataeopsephus japonicus Lampyridae Luciola cruciata Acari - Acarina N/A N/A 11

12 Table S2. Numbers of individuals for each taxon (the first sample replication) shown as the value per quadrat (30 cm 30 cm area) for each site. #

13 Table S2. Continued. #

14 Table S2. Continued. #

15 Table S2. Continued. #

16 Table S2. Continued. #

17 Table S2. Continued. #

18 Table S2. Continued. #

19 Table S3. Numbers of individuals for each taxon (the second sample replication) shown as the value per quadrat (30 cm 30 cm area) for each site. #

20 Table S3. Continued. #

21 Table S3. Continued. #

22 Table S3. Continued. #

23 Table S3. Continued. #

24 Table S3. Continued. #

25 Table S3. Continued. #