Trait characteristics determine pyrethroid sensitivity in non-standard test species of freshwater

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1 Supporting Information Trait characteristics determine pyrethroid sensitivity in non-standard test species of freshwater macroinvertebrates A reality check Peter Wiberg-Larsen, Daniel Graeber, Esben Astrup Kristensen, Annette Baattrup-Pedersen, Nikolai Friberg, Jes Jessen Rasmussen * Institute for Bioscience, Aarhus University, Vejlsoevej 25, DK-8600 Silkeborg, Denmark NIVA, Section for Freshwater Biology, Gaustadalléen 21, NO-0349 Oslo, Norway * Corresponding author: Jes Jessen Rasmussen jr@bios.au.dk The supporting material contains 6 tables and 5 figures: Table S1: Taxonomic details for test animals Table S2: Overview of sampling sites for test animals Table S3: Chemical composition of AFW Table S4: Overview of the mathematical models used to calculate volumes of test animals Table S5: Quantified lambda-cyhalothrin concentrations Table S6: LC 50 concentrations and control mortality Figure S1: Photos of the video tracking equipment Figure S2: LC 50 as a function of LOEC behavior Figure S3: PCA of species specific trait profiles Figure S4: Eureqa test details Figure S5: Correlations of LC 50 to species indicator scores in traditional ecological indices S1

2 Table S1. List of the taxonomic details for the macroinvertebrates used for toxicity testing. Order Family Genus / species Tricladia Dugesiidae Dugesia gonocephala Oligochaeta (subclass) Tubificidae Hirudinea (subclass) Glossiphonidae Helobdella stagnalis Hirudinea (subclass) Erpobdellidae Erpobdella octaculata Amphipoda Gammaridae Gammarus pulex Isopoda Asellidae Asellus aquaticus Plecoptera Nemouridae Protonemoura meyeri Plecoptera Nemouridae Nemoura cinerea Plecoptera Perlodidae Isoperla grammatica Plecoptera Leuctridae Leuctra fusca Ephemeroptera Caenidae Caenis horaria Ephemeroptera Baetidae Cloeon inscriptum Ephemeroptera Heptagenidae Heptagenia sulphurea Ephemeroptera Ephemeridae Ephemera danica Megaloptera Sialidae Sialis lutaria Coleoptera Dytiscidae Oreodytes sanmarkii Coleoptera Scirtidae Elodes minuta Coleoptera Elmidae Elmis aenea Coleoptera Elmidae Limnius volckmari Trichoptera Glossosomatidae Agapetus fuscipes Trichoptera Hydropsychidae Hydropsyche angustipennis Trichoptera Limnephilidae Chaetopteryx villosa Trichoptera Goeridae Silo pallipes Trichoptera Beraeidae Beraeodes minutus Diptera Ptychopteridae Ptychoptera paludosa Diptera Pediciidae Dicranota Diptera Simuliidae Odagmia ornata Diptera Chironomidae Conchapelopia menalops Diptera Chironomidae Prodiamesa olivacea Diptera Chironomidae Tvetenia Diptera Chironomidae Chironomus luridus agg. Diptera Syrphidae Eristalis Gastropoda Lymnaeidae Radix balthica Bivalvia (class) Sphaeriidae Sphaerium corneum S2

3 Table S2. Overview of sampling sites for the 34 macroinvertebrate species/taxa. Species/genus/family Sampling site ID / type J: Jutland F: Fyn Distribution in Denmark: P -prevalent /abundant L - local Sampling time (month/year) Behavioural assessment using video tracking Dugesia gonocephala (TC) Hoejbjerg brook (F)/stream P (Jutland/Fyn) 10/2011 Yes Tubificidae (O) Holme Brook (F)/stream P 3/2012 No Helobdella stagnalis (H) Assenboelle Brook P 8/2012 No (F)/stream Erpobdella octaculata (H) Hoejris Aa (J)/stream P 9/2011 No Gammarus pulex (CA) Lindved Aa (F)/stream P 10/2005 Yes Asellus aquaticus (CI) Traunskov (F)/pond P 9/2011 Yes Protonemoura meyeri (P) Jeksen Brook (J)/stream L (Jutland) 3/2012 Yes Nemoura cinerea (P) Vrads Brook (J)/stream P 3/2012 Yes Isoperla grammatica (P) Jeksen Bæk (J)/stream P/L 4/2012 Yes Leuctra fusca (P) Stavis Aa (F)/stream P 6/2012 Yes Caenis horaria (E) Vestermose (F)/pond P 4/2012 Yes Cloeon inscriptum (E) Elverod (F)/pond P 4/2012 Yes Heptagenia sulphurea (E) Brende Aa (F)/stream P 4/2012 Yes Ephemera Danica (E) Gudenaa (J)/stream P 11/2011 Yes Sialis lutaria (M) Brorenden (F)/stream P 11/2011 Yes Oreodytes sanmarkii (CO) Skaerup Aa (J)/stream P (Jutland) 6/2012 Yes Elodes minuta (CO) Brorenden (F)/stream P 11/2011 No Elmis aenea (CO) Kragelund Millbrook P 5/2012 No (F)/stream Limnius volckmari (CO) Stavis Aa (F)/stream P 5/2012 No Agapetus fuscipes (TR) Limkilde (F)/stream P/L 3/2012 No Hydropsyche angustipennis Milling Brook (J)/stream P 4/2012 Yes (TR) Chaetopteryx villosa (TR) Jeksen Brook (J)/stream P 4/2012 Yes Silo pallipes (TR) Jeksen Brook (J)/stream P 4/2012 Yes Beraeodes minutus (TR) Brorenden (F)/stream L 11/2011 No Ptychoptera paludosa (D) Gudenaa (J)/stream P 11/2011 No Dicranota (D) Hadsten Lilleaa (J)/stream P 3/2012 Yes Odagmia ornata (D) Vrads Brook (J)/stream P 3/2012 No Conchapelopia menalops (D) Lystrup Aa (J)/stream P 5/2012 No Prodiamesa olivacea (D) Salten Aa (J)/stream P/L 4/2012 No Tvetenia (D) Ravning Forest (J)/stream P 8/2012 No Chironomus luridus agg. (D) Kohave (F)/stream L 8/2012 No Eristalis (D) Hedemose (J)/stream L 8/2012 No Micropsectra (D) Salten Aa (J)/stream P 8/2012 No Radix balthica (G) Hoejris Aa (J)/stream P 9/2011 No Sphaerium corneum (B) Stavis Aa (F)/stream P 8/2012 No Notes: TC Turbellaria, Tricladida; O Oligochaeta; H Hirudinea; CA Crustacea, Amphipoda; CI Crustacea, Isopoda; P Insecta, Plecoptera; E Insecta, Ephemeroptera; M Insecta, Megaloptera; CO Insecta, Coleoptera; TR Insecta, Trichoptera; D Insecta, Diptera; G Gastropoda; B Bivalvia. S3

4 Table S3. Chemical concentrations in the test medium. Salt Concentration (mg L -1 ) CaCl 2 2H 2 O 294 MgSO 4 7H 2 O NaHCO KCl 5.75 S4

5 Table S4. Overview of the three-dimensional models used to calculate surface areas of the test organisms. For select species, adjustment factors were used to adjust the surface area with respect to larger extremities such as antennae, cerci and gills. The following abbreviations are used: L = length; W = width; H = height; e= 1, where a = ½W and c = ½L. Species/taxa Shape type Mathematical model Adjustment factor a Dugesia gonocephala None None None Tubificidae None None None Helobdella stagnalis None None None Erpobdella octaculata None None None Gammarus pulex None None None Asellus aquaticus Ellipse (π 0.25 L W) ( L H) Protonemoura meyeri Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) 1.15 Nemoura cinerea Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) 1.10 Isoperla grammatica Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) 1.10 Leuctra fusca Cylinder 2π½WL +2π(½W) Caenis horaria Ellipse (π 0.25 L W) ( L H) Cloeon inscriptum Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) 1.20 Heptagenia sulphurea Triangle x height ½WL+2HL+HW 1.20 Ephemera danica Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) 1.20 Sialis lutaria Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) 1.15 Oreodytes sanmarkii Ellipse (π 0.25 L W) ( L H) Elodes minuta Ellipse (π 0.25 L W) ( L H) Elmis aenea (adult) Ellipse (π 0.25 L W) ( L H) Limnius volckmari (larva) Cylinder 2π½WL +2π(½W) Agapetus fuscipes Cylinder 2π½WL +2π(½W) Hydropsyche angustipennis Cylinder 2π½WL +2π(½W) Chaetopteryx villosa Cylinder 2π½WL +2π(½W) Silo pallipes Cylinder 2π½WL +2π(½W) Beraeodes minutus Cylinder 2π½WL +2π(½W) Ptychoptera paludosa Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) 1.10 Dicranota Cylinder 2π½WL +2π(½W) 2 None Odagmia ornata Spheroid prolate 2πa 2 (1+(c/ae)sin -1 e) None Conchapelopia menalops Cylinder 2π½WL +2π(½W) 2 None Prodiamesa olivacea Cylinder 2π½WL +2π(½W) 2 None Tvetenia Cylinder 2π½WL +2π(½W) 2 None Chironomus luridus agg. Cylinder 2π½WL +2π(½W) Eristalis Cylinder 2π½WL +2π(½W) Radix balthica None None None Sphaerium corneum None None None a The adjustment factor takes into account the increased surface are introduced by the presence of extremities, number of extremities, and size and form of these extremities. S5

6 Table S5. Verified exposure concentrations of lambda-cyhalothrin. Statistics Nominal lambda-cyhalothrin concentration (µg L -1 ) Mean Median SD SE CV N (no. analyses = taxa) S6

7 Table S6. LC 50 concentrations and mortality in untreated controls for the tested species. LC 50 concentrations are given ± SE. In the cases where no mortality was observed, the test species was ascribed an artificial LC 50 concentration of 1,000 µg L -1. Test species LC 50 (µg L -1 ) Control mortality (%) Agapetus fuscipes ± Asellus aquaticus ± Beraeodes minutus 3.75 ± Caenis horaria 0.14 ± Chaetoptyeryx villosa 0.79 ± Chironomus lugubris agg ± Cloeon inscriptum 1.56 ± Conchapelopia menalops ± Dicranota sp Dugesia Gonocephala Elmis aenea 0.41 ± Elodes minuta 1.26 ± Ephemera danica 9.35 ± Eristalis Erpobdella octaculata Gammarus pulex 0.46 ± Helobdella stagnalis Heptagenia sulphurea 0.88 ± Hydropsyche angustipennis Isoperla grammatica ± Leuctra fusca 1.43 ± Limnius volckmari 3.24 ± Micropsectra 0.75 ± Nemoura cinerea 0.48 ± Odagmia ornata 0.14 ± Oreodytes sanmarkii 8.79 ± Prodiamesa olivacea Protonemoura meyeri ± Ptychoptera paludosa 1.4 ± Radix baltica Sialis lutaria Silo pallipes 1.25 ± Sphaerium corneum Tubificidae Tvetenia 0.63 ± S7

8 Figure S1. Experimental set up of the EthoVision videotracking system to the left and a camera view of the tracking system to the right. S8

9 Fig. S2. LC 50 concentrations as a function of the LOEC for behavioral responses during lambdacyhalothrin exposure. The exposure duration was 90 min, and lethality was recorded 6 days postexposure. The red dashed line indicates the 1:1 relationship. The abbreviations for species names represent Leuctra fusca, Protonemoura meyeri, Cloeon inscriptum, Heptagenia sulphurea, Gammarus pulex, Asellus aquaticus, Isoperla grammatica, Nemoura cinerea, Caenis horaria, Ephemera Danica, Silo pallipes, and Chaeopteryx villosa. S9

10 1.0 Surface area/weight Size > cm PCA 2 (21.5%) Size > cm Tegument respiration Medium flow Macrophytes Fast flow Boulder Surface area Twigs/roots Gravel Slow flow Size > 2-4 cm Null flow Size > 1-2 cm Weight Silt Mud Sand Organic detritus -1.5 Gill respiration PCA 1 (27%) Fig. S3. Principal Component Analysis of the species specific trait profiles. S10

11 Fig. S4. Predictive ability parsimony Pareto front of model results (a.) and number of models each independent variable occurs in (b.). In panel a., the selected model is marked with a red dot. Variables, which appeared in none of the models, are not shown in panel b. S11

12 Fig. S5. Log LC 50 concentrations as a function of the indicator values for the test species in the DSFI (A), BMWP (B) and the SI (C) indices, respectively. S12

APPENDIX B THREE RIVERS SECOND NATURE RESULTS OF RAPID INVERTEBRATE BIOLOGICAL ASSESSMENT SAMPLING PHASE 2 APRIL 2002 A-19

APPENDIX B THREE RIVERS SECOND NATURE RESULTS OF RAPID INVERTEBRATE BIOLOGICAL ASSESSMENT SAMPLING PHASE 2 APRIL 2002 A-19 Shades Run Squaw Run Sandy Quigley Indian Powers Run Phylum Arthropoda Insecta