Three replicate permanent transects (10 × 1 m) on each reef patch were also established within the immediate vicinity of decomposing A. planci. These transects were very small relative to normal sampling protocols for coral reef fishes, but this was sufficient given the very small area of impact – all decomposing A. planci were within an area measuring approximately 10 m × 6 m. Injected A. planci were mostly hyperactive up to an hour after injection,

LDK378 but subsequently remain stationary prior to death. A video recording (distance of 0.5 m from the substrate) of the entire length of each permanent transect was done on day 1, day 7, and day 14 to monitor fish and macro-invertebrate populations. In addition, 20 colonies of branching corals (Acropora, Pocillopora, Seriatopora and Stylophora) were individually tagged and then photographed at regular intervals (every 1–5 days) to test for any new incidences Lapatinib supplier of coral disease. These colonies were located at distances of 0–4 m from the A. planci aggregations. The main parameters analyzed are mortality (proportion of individuals dead after 48 h) and time until death after injection (Table S1). These are important factors in assessing the efficiency of any method of killing COTS and its feasibility as a control measure. Differences in mortality (proportion dead after 48 h) between bile derivatives, dosage, and sites of injection were

compared using Fisher’s Exact Test (Table S2; Sokal and Rohlf, 1995). Time until death in COTS injected with bile salts and oxgall at different concentrations (Table S3) were also analyzed by performing a two-way Scheirer-Ray-Hare extension of the Kruskal–Wallis test (Table S4; Sokal and Rohlf, 1995). Only those injected in the central disk was included in this analysis because double

dose treatments were only injected on this part of the seastar. Variation in time to death after injection Galeterone between the two bile derivatives tested and the four sites of injection (Table S5) were analyzed using Two-way (Model II) ANOVA, with both parameters as random factors (Table S6; Sokal and Rohlf, 1995). COTS that recovered and survived after the 7-day observation period was assigned a time of 168 h in order have equal sample sizes for each treatment. Data were log-transformed prior to analysis to satisfy assumptions of normality and homogeneity of variance. All COTS injected with Bile Salts No. 3 experienced 100% mortality regardless of the concentration and site of injection (Fig. 1A, C). This was significantly higher compared to sea stars injected with Oxgall (Fig. 1B, D), which only experienced 80% mortality after 48 h (p = 0.022, Table S2). There was no significant increase in mortality even when concentrations of bile derivatives were doubled (p = 1.000, Table S2). Among the COTS injected in the central disk ( Table S3), those injected with Bile Salts No. 3 (27.90 ± 6.84) died more rapidly (H1,16 = 4.117, p = 0.