1. Effect of Neem (Azadirachta indica) Seed Extract on Artemia Alexandra Mendez, Yadhira D. Arocho, Lizza Rodriguez, Christopher Cardona, and Eythan Morenu. University of Puerto Rico at Aguadilla

2. Effect of Neem (Azadirachta indica) Seed Extract on Artemia By: Alexandra Mendez, Yadhira D. Arocho, Lizza Rodriguez, Christopher Cardona, and Eythan Morenu. Evaluation of data and conclusions Quick View of Artemia organism Quick View of Neem tree Experimental process

3. Phylum Arthropoda Subphylum Crustacea Class Branchiopoda Order Anostraca Family Artemidae Genus Artemia Clasification of the brine shrimp Artemia:

4. oviparous ovoviviparous Life cycle of Artemia Continuous reproduction Alternation of reproductive mode Production of encysted embryos Production of nauplii There are sexual and parthenogenetic species

5. Artemia is a very important organism from an ecological and economic perspective Artemia biomass is used in aquaculture (fish food), this is a million dollar industry Artemia are important for the production of salt and the survival of many species of birds in the salt flat system

6. Research question: How will the Neem (Azaridachta indica) seed extract affect the survival of Artemia larvae?

7. Neem Tree Info. Escribir informacion relacionada al arbol de NEEM. !

8. Neem = Insecticide Effect of Neem in Artemia = ? ?

9. 50% 35% 10% 5% 0% (CONTROL) From the original filtered stock solution of 200 g/L: Prepared Solutions

10. 100% 50% 40% 20% 10% From the original filtered stock solution of 200 g/L: 5% 0% = CONTROL

11. Artemia cysts from The Great Salt Lake Neem seeds, collected from a tree located in Cabo Rojo, P.R., –The seeds were placed in a food processor with 100 mL of off-shore sea water with a salinity of 35. – The obtained solution was filtered twice through a 0.5mm filter. Experimental Process

12. 10 Artemia nauplii for each of the tubes 25 test tubes Total of 250 Artemia nauplii

13. The experimental test tubes containing the Artemia nauplii were placed in a test tube holder, and were left undisturbed in a dark laboratory room with a temperature of 25°C until the next day.

14. 24 hours later hours later (the next day) the experimental test tubes were observed and the live and/or dead Artemia nauplii were counted.

15. Both experimental phases were based on a filtered stock neem seed solution of 200g/L. The difference between both experimental phases was the concentration of the solution used on the experiment but both phases parted from the original 200g/L stock solution. In phase one we did not find significant death rates on Artemia at the different concentrations, this was our main motivation to repeat the experiment (phase II).

16. In phase II we modified a little the concentrations (modifications shown on methods and materials section) and implemented a 100% concentration. We also implemented a dissolved oxygen test performed with a Hach HQ10 dissolved oxygen meter.

17. Results?

20. Our results for phase I: In phase one we did not find significant death rates on Artemia at the different concentrations, this was our main motivation to repeat the experiment (phase II).

21. Our results for phase II: Lethal effect on Artemia Artemia died at the different concentrations. However, we can not conclude that Artemia died because of the neem seed extract because our dissolved oxygen tests demonstrated levels of dissolved oxygen that were too low for the Artemia organism (dissolved O2 levels shown on graph 2).

22. On our next experimental effort (phase III) we will perform the experiment with a controlled oxygen environment that is adequate for the Artemia and this will enable us to determine the effect neem seed extract might have on Artemia on the different concentrations.

23. In an aquaculture practical application we want to know if neem seed extract has a positive effect on the growth and development of Artemia. If neem seed extract would promote a faster growth rate it could be used to accelerate development of Artemia and more Artemia could be grown faster to satisfy the growing demand of Artemia in the aquaculture industry.

24. Acknowledgements Profa. Sandra Quiñones Prof. Carlos Ruiz Profa. Liza Jimenez Profa. Sonia Rivera Profa. Migdalia Sotomayor Prof. Robert J. Mayer Prof. Rafael Estremera