COTS were placed in individual 68-l plastic containers with flow-through seawater at ambient conditions. Injections of 10 ml of each solution (initially, 4 g l−1 of Bile Salts No. 3 and 6 g l−1 of Oxgall) were administered using a plastic

syringe with an 18-gauge needle. Sea stars were injected in (1) the distal portion of the arm, (2) the middle of the Ganetespib manufacturer arm, (3) the proximal portion of the arm, and (4) the central disk ( Fig. 1). A. planci used in the double dose treatments were all injected in the central disk. Two separate measures of the effectiveness are considered in this study: i) the time until death (in hours), recorded as the time from injection until all podia (tube feet) were completely immobile ( Rivera-Posada et al., 2011), and ii) the proportion of sea stars that actually died with 2–3 days. A total of 12 A. planci distributed in three groups of 4 sea stars were used for this experiment. Each A. planci was injected with 10 mL of 8 g l−1 Bile Salts No. 3 and time to death was estimated.

Hyperactivity shortly after injection was used as an indicator that the sea star was correctly injected. Three different types of injection guns were tested ( Fig. 2): (1) DuPont™ Velpar® Spotgun®, (2) Simcro™ STV 12-ml Plastic Syringe, and (3) prototype metal injection gun. The DuPont™ Velpar® Spotgun® was fitted with a 50-cm needle, 4-mm tip, and 5-L plastic bladder, which is currently used in the field to inject sodium bisulfate (dry acid) solution. Although this gun provides good reach to cryptic A. planci, the INCB024360 research buy width of the tip creates large holes, which raises concerns that chemicals injected could easily leak out of these openings without any effect or without killing the sea star. It is important to note that this gun was originally designed to spray herbicides and not to inject A. planci. Simcro™ STV 12-ml Plastic Syringe is cheap, lightweight, requires minimal maintenance, and offers the possibility to attach Montelukast Sodium any size and length of syringe needle. This gun has been successfully used in A. planci control efforts around Japan ( Kuroshio Biological Research

Foundation, 2011). A prototype metal injection gun with a 50-cm spear and Luer-lock to attach a 16 Ga × 1/2″ needle was designed for more accurate injections of small amounts of solution ( Fig. 2, inset). A thinner and shorter needle was used to minimize the puncture size and leakage after injection and to avoid overshooting (tip of needle exits the sea star arm and solution is not injected internally) during injection, as what usually happens with longer needles. Fish, corals, and other echinoderms (Table 1) were collected from back reef habitats around Lizard Island. Smaller fishes (i.e. Pomacentridae, Chaetodontidae) were collected using clove oil, which is noteworthy because clove of its hepatotoxic properties (Javahery et al.