1. Semiochemicals as Shark Repellents: Identification and Behavioral Response Eric M. Stroud Michael M. Herrmann Dr. Samuel G. Gruber

2. Semiochemical Mixture Semiochemicals are messengers or “clues” –Shark-specificity –Non-lethal –Action is suspected to be olfactory (mouth dose is not required) –Not based on Cu+, surfactants, acid, or dye –Safe to handle and store

3. Discovery of the Mixture Shark carcass – Not decayed, no Cu+, no ammonium acetate Production –Multi-phase extraction process –Deproteinization –Filtration –Neutralization Modern analytical instrumentation Mixture of nucleotides

4. Analytical Tools Visible / IR Spectrophotometry Fluorimetry Derivatization and HPLC Flash Chromatography and TLC GC-MS

5. GC-MS Total Ion Chromatograph

6. Field Tests Find a suitable wild shark population Construct a delivery apparatus Test a behavioral control Test a variety of fractions of the semiochemical mixture Quantify results

7. Testing Locations BBFS – South Bimini –Blacknose, Caribbean Reef, Lemon, Bull sharks Ordinance Dump – Montauk, NY –Blue sharks

8. Delivery Apparatus Based on Donald Nelson’s design Pressurized delivery of liquid through a tube to a fish head suspended on a pole Pressures ranges from 5- 20 psig Delivery volumes ranges from 100mL to 500mL

9. Controls and Repellents Control is introduced while sharks are actively feeding –Seawater with or without FD&C Red 3 & 40, at the same pressure and dose size for the repellent Repellent is introduced while sharks are actively feeding –Observations are made on the size of the feeding population before and after dosing –Observations are made on changes in feeding behavior

10. Results Controls –Feeding shark populations were not diminished for control deliveries of up to 20 psig –Plain seawater nor FD&C Red 3 &40 and seawater did not deter feeding –Sharks are not afraid to feed from and near the delivery apparatus

11. Results Repellents –100mL to 250mL doses of the semiochemical mixture dramatically reduce the feeding population size –Nearby fish are unaffected and continue feeding –In six separate field tests, feeding populations of 2 to 12 Blacknose and Caribbean Reef sharks are reduced to 0 within 120 seconds of a subsurface dose –Visual behavior changes occur within 20 seconds of the subsurface dose –4 fl oz is the optimal dose size for large populations, but response has been observed with as little as 25mL

12. Species and Populations Blacknose / Caribbean Reef Sharks – Up to 12 Blue Sharks – 2 Bull Sharks – 2 Lemon Sharks – 2 Nurse Sharks - 2

13. Cloud Test: Population = 12

14. Tonic Immobility Test

15. 0.1ppm Criteria Test for a behavioral response at with 0.1 ppm of the chemical repellent Johnson-Baldridge proposed that a 100mL/hr release of a chemical repellent into a 6m 3 boundary of water under steady-state conditions for 3.5 hours represents 0.1 ppm.

16. 0.1ppm Criteria Use a tall plastic tripod with wireless camera to observe a 6m 3 boundary of water, with a fish head suspended in the 6m 3 boundary Chum to attract sharks into the boundary area, let them take the fish head Begin metered dosing of the repellent using a precision peristaltic pump into the boundary once a population is established, using a new fish head New fish head must remain protected for 3.5 hours

17. Johnson-Baldridge Apparatus

18. 0.1ppm Results September 2003 –Fish head was protected for 1.5 hours from two lemon sharks, until medical pump battery died March 2004 –Fish head was protected for 2 hours from two Caribbean reef sharks, until pump failed May 2004 –Successfully performed and video’d a control

19. Next Steps More field testing: –Tiger, Great White, Hammerhead, Oceanic Whitetip More validation: –Bull, blue Perfect synthesis pathway for commercialization

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