<p>Supplementary Information: Climate change as an unexpected co-factor promoting coral eating seastar (Acanthaster planci) outbreaks</p><p>S. Uthicke, M. Logan, M. Liddy, D. Francis, N. Hardy, M. Lamare Day 10 ) ) )</p><p>% 60 % %</p><p>15 (</p><p>( ( 80 .</p><p> l h a a i c 10 r 40 a a m 60 r r n o B n</p><p> i n</p><p> y 20 p</p><p>5 l b 40 i r A B a</p><p>0 20 E 0 ) ) 25 10 )</p><p>% 1 1.5 % ( l − (</p><p>8 . 20 . m h h (</p><p> c c 6 1.0</p><p>15 y a a t r i r B s B 4</p><p>10</p><p> n 0.5 d e i e 5 t 2 a D M 0 L 0 0.0 2 6 10 2 6 10 2 6 10 Day 17 ) ) )</p><p>35 60 % ( %</p><p>% 60</p><p>( . ( 30</p><p> l h</p><p> a 50 i c a 25 50 r a a m r</p><p> r 20 n 40 40 B o</p><p> n i n</p><p>15 y l p b r 10 i 30 30 a A B 5 20 E 20 ) ) 8 1.4 ) % 1 %</p><p>( 25 ( l − 1.2</p><p>. . 6 m h h 20 ( 1.0</p><p> c c y a a t</p><p> r 15 0.8 i r 4 B s B</p><p>10 n 0.6 d e i e t 2</p><p> a 0.4 D</p><p>M 5 0 L 0 0.2 2 6 10 2 6 10 2 6 10 Day 24 ) ) 40 ) 60</p><p>% 60 % % (</p><p>( ( . 50 50 l</p><p>30 h a a i c r 40 40 a a m r r 20 n 30 o</p><p>30 B n</p><p> i n</p><p> y 20 p l b</p><p> i 20</p><p>10 r A B a 10 10 0 E 0 ) ) )</p><p>% 60 1 %</p><p>( 40 ( − l</p><p>.</p><p>. 50 m h h 30 (</p><p> c</p><p> c 40 y a a t r i r 30</p><p>B 20 s B</p><p> n</p><p> d 20 e i e 10 t a D</p><p>M 10 0 L 0 0.0 2 6 10 2 6 10 2 6 10 Algae cells (x 1000 ml−1)</p><p>Supplementary Figure 1: The percentage of abnormal larvae (Abnormal), Bipinnaria, Early Brachiolaria, Mid Brachiolaria, Late Brachiolaria and larval densities of Acanthaster planci at 3 days during the experiments, different colours indicate temperature treatments (Blue = 28°C, Green = 29°C and Red = 30°C). Please note that some symbols are hidden behind others. ) 600 ) 700 m m µ</p><p>550 µ ( ( 600</p><p> h t h 500 t g d i 450 n 500 e W L</p><p> y 400 y</p><p> d 400 d</p><p> o 350 o B 300 B 300 28 30 28 29 30 Temperature( °C) 200 ) ) 140 m m µ µ (</p><p>( 120</p><p>150 h h t t 100 g d i n e 100 W</p><p>80 L t</p><p> t u u</p><p>G 60 G 50</p><p>1.1 4.2 9.8 1.1 4.2 9.8</p><p>Algae cells (x 1000 ml-1)</p><p>Supplementary Figure 2: Biometric measures showing some degree of correlation with Temperature (top two panels) or food concentration (bottom two panels) in the redundancy analysis of D10. Box and whisker plots are based on average values per replicated (experimental container). Whiskers denote 1.5 x the inter-quartile range, the black line indicates the mean. ) 900 ) 700 m m µ µ ( 800 ( 600</p><p> h h t</p><p>700 t g</p><p> d 500 i n 600 e W</p><p>L 400 500 y y d d</p><p>400 o 300 o B B )</p><p>400 ) 400 m m µ</p><p>( 350 350 µ</p><p>(</p><p> d 300 300 d o o o</p><p>250 o 250 H</p><p>H</p><p> h</p><p>200 t 200 t u u</p><p> o 150 150 G M ) 200 ) 300 m m µ µ ( 180 ( 250</p><p> h h t 160 t d i</p><p> g 200</p><p>140 n W e 150 L h 120</p><p> t t u 100 u 100 o G M 1.1 2.8 4.2 9.8</p><p>) 200 m µ (</p><p>150 h t d i 100 W</p><p> t u 50 G 1.1 2.8 4.2 7.0 9.8</p><p>Algae cells (x 1000 ml-1)</p><p>Supplementary Figure 3: The relationship of food concentrations on seven biometric measures of Acanthaster planci larvae at D24. All measures also showed some degree of correlation to food concentrations in the redundancy analysis. Box and whisker plots are based on average values per replicated (experimental container). Whiskers denote 1.5 x the inter-quartile range, the black line indicates the mean.</p><p>31.0</p><p>30.5</p><p>30.0 )</p><p>C 29.5 ° ( e r u</p><p> t 29.0 a r e p 28.5 m e T 28.0</p><p>27.5</p><p>27.0</p><p>26.5</p><p>26.0 17/11 22/11 27/11 2/12 7/12 12/12 Date (</p><p>Supplementary Figure 4: Temperature profiles for the three temperature treatments referred to as 28°C (average: 27.83°C, SD = 0.13°C), 29°C (average: 28.75°C, SD = 0.05°C) and 30°C (average: 29.81°C, SD = 0.05°C). Temperatures were recorded every minute (N = 39399) using Hoboware temperature loggers. Supplementary Table 1. Details of the slopes and intercepts of generalized linear models testing the effects of food concentrations and temperature on the percentage of Acanthaster planci larvae developed to mid and late brachiolaria stage at three measurement days. </p><p>Estimate SE t p</p><p>D 10 Intercept -4.704 0.587 -8.01 <0.0001 Algae 0.179 0.072 2.50 0.0152 Temperature 29 0.869 0.519 1.68 0.0993 Temperature 30 1.637 0.457 3.58 0.0001 D 17 Intercept -3.580 0.280 -12.80 <0.0001 Algae 0.241 0.037 6.52 < 0.0001 Temperature 29 -1.088 0.617 -1.76 0.0845 Temperature 30 -0.957 0.653 -1.47 0.1494 Algae x Temperature 29 0.232 0.098 2.37 0.0219 Algae x Temperature 30 0.280 0.110 2.54 0.0146 D 10 Intercept -2.054 0.359 -5.72 <0.0001 Algae 0.268 0.053 5.07 < 0.0001 Temperature 29 -0.315 0.359 -0.88 0.3850 Temperature 30 0.075 0.400 0.19 0.8520 Supplementary Table 2 Summary of additional observations during the feeding and </p><p> temperature experiment on Acanthaster planci at each of the temperature and algae treament </p><p> combinations. Un-induced settlement refers to settlement and metamorphosis in the </p><p> experimetnal containers. Induced settlement refers to a trial on D24, offering Lithothamnium </p><p> pseudosorum in 6 well containers . + and – refers to the presence or absence of </p><p> metamorphosed larvae after 24h. Lithothamnium pseudosorum is the preferred settlement </p><p> substrate for A. planci larvae </p><p>Temperature Algae First Late Brachiolaria First un-induced settlement Induced settlement (°C) (cells ml-1) (days) (days) (D 24) 28 1100 24 >35 - 2800 24 >35 - 4200 17 24 + 7000 17 20 + 9800 10 22 + 29 1100 24 >35 - 4200 17 24 + 7000 17 20 + 30 1100 24 >35 - 4200 10 20 + 7000 10 20 + Supplementary Table 3: Algae concentrations (in cell numbers ml-1), consisting of 8% </p><p>Dunialiella, 62% Phaeodactylum and 31% Chaetocerus, and measured chlorophyll concentrations in the treatment containers. Given is the average of 4 measuring days, the standard deviation is given in brackets. The average chlorophyll concentration is significantly related to the target number of algae (R2 = 0.99, p < 0.001).</p><p>Algae Chlorophyll (µg l-1) 1100 0.67 (0.31) 2800 1.80 (0.90) 4200 2.55 (1.07) 7000 4.85 (1.33) 9800 7.11 (1.18)</p><p>Supplementary references</p><p>Johnson, C. R., D. C. Sutton, R. R. Olson, and R. Giddings. 1991. Settlement of crown-of-thorns starfish: role of bacteria on surfaces of coralline algae and a hypothesis for deepwater recruitment. Mar. Ecol. Prog. Ser. 71:143-162. Uthicke, S., D. Pecorino, R. Albright, A. P. Negri, N. Cantin, M. Liddy, S. Dworjanyn, P. Kamya, M. Byrne, and M. Lamare. 2013. Impacts of Ocean Acidification on Early Life-History Stages and Settlement of the Coral-Eating Sea Star Acanthaster planci. PLoS ONE 8:e82938.</p>