Testing the Neem Biopesticide (Azadirachta indica A. Juss) for Acute Toxicity with Danio rerio and for Chronic Toxicity with Daphnia magna

Authors
L. A. Maranho 1
R. G. Botelho 1
M. Mitie Inafuku 1
L. D. A. R. Nogueira 1
R. Alves de Olinda 2
B. A. Inacio de Sousa 1
V. L. Tornisielo 1
1 Center for Nuclear Energy in Agriculture, University of São Paulo - CENA/USP, Laboratory of Aquatic Toxicology, Av. Centenario, 303, São Dimas, CEP: 13416-000, - Piracicaba, SP, Brazil.
2 Estadual University of Paraíba - UEPB. Campus I - Center for Science and Technology, Department of Statistic. Av das Baraúnas, 351, CEP 58101-001, Bodocongó, Campina Grande, Paraíba, Brazil.
Abstract
Recently, some natural products have been used in the fields as alternative to synthetic compounds, to minimize the negative impacts to the environment. This study aimed to verify the effects of Neem-based bio-pesticide in causing acute toxicity for a fish and chronic toxicity for a microcrustacean. To this end, Danio rerio and Daphnia magna were exposed to various concentrations of a Neem-based oil formulation. In the first experiment, adults of D. rerio were exposed for 96 hours to different concentrations to determine the median lethal concentration (LC50-96h). For D. magna, first an acute toxicity test was performed to determine the median effective concentration (EC50-48h). Based on the EC50 established in the acute test, the concentrations for the 21-day chronic toxicity test were determined. Endpoints evaluated were reproduction (number of neonates produced) and size of D. magna.The median lethal concentration for the fish was 0.22 mL L-1, and the median effective concentration (EC50 - 48h) for D. magna was 0.17 mL L-1. In the chronic test, all concentrations affected reproduction and size of D. magna. The formulation tested may be hazardous to aquatic organisms.

Keywords

1. ABNT. 2003. NBR 12713: Norma Técnica: Ecotoxicologia Aquática– Toxicidade Aaguda-Método de Eensaio com Daphnia spp (Crustacea- Cladocera). ABNT, Rio de Janeiro, Brazil, 17pp.
2. Ahmad, M. K. and Ansari, B. A. 2011. Toxicity of Neem-based Pesticide Aazacel to the Embryo and Fingerlings of Zebrafish Danio rerio. W. J. Zoology, 6: 47-51.
3. Botelho, R. G., Inafuku, M. M, Maranho, L. A, Machado Neto, L, Olinda, R. A, Dias, C. T. and Tornisielo, V. L. 2010. Toxicidade Aguda e Crônica do Eextrato de Nim (Azadirachtas indica) Para Ceriodaphnia dúbia. Pesticidas: R. Ecotox. Meio. Embiente, 20: 29-34.
4. Box, G. E. P. and Cox, D. R. 1964. An Analysis of Transformations (with Discussion). J. R. Stat. Soc. Series B., 26: 211-252.
5. Carneiro, M. T. P. 2003. Efeito de Extratos de Folhas e do Oleo de Nim Ssobre o oídio do Tomateiro. Summa Phytopathol., 29: 262-265.
6. Dunkel, F. V. and Richards, D. C. 1998. Effect of an Azadirachtin Formulation on Six Nontarget Aquatic Macroinvertebrates. Environ. Entomol., 28: 667-674.
7. Gandhi, M., Lal, R, Sankaranayanan, A, Banerjee, C. K. and Sharma, P. L. 1988. Acute Toxicity Study of the Oil from Azadirachta indica Seed (Neem Oil). J. Ethnopharmacol., 23: 39–51.
8. Goktepe, I. and Plhak, L. C. 2002. Comparative Toxicity of Two Azadirachtin-Based Neem Pesticides to Daphnia pulex. Environ. Toxicol. Chem., 21: 31–36.
9. Hamilton, M. A., Russo R. C. and Thurston, R. V. 1977. Trimmed Spearman-Karber Method for Estimating Median Lethal Concentrations in Toxicity Bioassays. Environ. Sci. Technol., 11: 714-719.
10. Hartley, H. O. 1950. Use of Range in Analysis of Variance. Biometrika, 37: 271–280.
11. Immich, A. P., Souza, A. A. U. and Souza, S. M. U. 2009. Removal of Remazol Blue RR dye from Aqueous Solutions with Neem Leaves and Evaluation of Their Acute Toxicity with Daphnia magna. J. Hazard. Mater., 164: 1580-1585.
12. Koul, O., Isman, M. B. and Ketka, C. M. 1990. Properties and Uses of Neem, Azadirachta indica. Can. J. Botany, 68: 1–11.
13. Mahboob, M., Siddiqui, M. J. K. and Jamil, K. 1998. The Effect of Subacute Administration of a Neem Pesticide on Rat Metabolic Enzymes. J. Environ. Sci. Health, Part B: Pestic., Food Contam. Agric. Wastes, 33: 425–438.
14. Mondal, D., Barat, S. and Mukhopadhyay, M. K. 2007. Toxicity of Neem Pesticides on a Fresh Water Loach, Lepidocephalichthys guntea (Hamilton Buchanan) of Darjeeling District in West Bengal. J. Environ. Biol., 28: 119-122.
15. Organization for Economic Co-operation and Development (OECD). 1992. Fish, Acute Toxicity Test: Effects on Biotic Systems. 203 Guidelines for Testing of Chemicals. OECD, Paris, 9pp.
16. Osuala, F. O. U. and Okwuosa, V. N. 1993. Toxicity of Azadirachta indica to Freshwater Snails and Fish, with Reference to the Physicochemical Factor Effect on Potency. Appl. Parasitol., 34: 63–68.
17. Santos, A. C. G., Rodruigues, O. G, Araújo, L. V. C, Santos, S. B, Guerra, R. M. S. N. C, Feitosa, M. L. T, Teixeira, W. C. and Santos-Ribeiro, A. 2006. Uso de Extrato de Nim no Controle de Acaríase por Myobia musculi Schranck (Acari: Miobidae) e Myocoptes musculinus Koch (Acari: Listrophoridae) em Camundongos (Mus musculus var. albina L.). Neotrop. Entomol., 35: 269-272.
18. Saucke, V. H. and Schumutterer, H. 1992. Untersuchungen uber Nebenwirkungen Verschiedener Niemprodukte bei Daphnia magna Strauss (Crustacea: Cladocera). Anz. Schädlingskd. Pfl. Umwelt., 65: 121–126.
19. Schaaf, O., Jarvis, A. P, Van Der Esch, S. A., Giagnacovo, G. and Oldhan, N. J. 2000. Rapid and Sensitive Analysis of Azadirachtin and Related Triterpenoids from Neem (Azadirachta indica) by High-performance Liquid Chromatography-Atmospheric Pressure Chemical Ionization Mass Spectrometry. J. Chromat. A., 886: 89-97.
20. Scott, I. M. and Kaushik, N. K. 1998. The Toxicity of Margosan-O, a Product of Neem Seeds, to Select Target and Nontarget Aquatic Invertebrates. Arch. Environ. Contam. Toxicol., 35: 426-431.
21. Schmutterer, H. 1990. Properties and Potential of Natural Pesticides from the Neen Tree, Azadirachta indica. Annu. Rev. Entomol., 35: 271-297.
22. Stark, J. D. and Walter, J. F. 1995. Persistence of Azadirachtin A and B in Soil: Effects of Temperature and Microbial Activity. J. Environ. Sci. Health, Part B: Pestic., Food Contam. Agric. Wastes., 30: 685–698.
23. Wan, M. T., Watts, R. G, Isman, M. G. and Strub, R. 1996. Evaluation of Acute Toxicity to Juvenile Pacific North West Salmon of Azadirachtin, Neem Extract, and Neem Based Products. Bull. Environ. Contam. Toxicol., 56: 432 – 439.
Journal of Agricultural Science and Technology
Volume 16, Issue 1
January and February 2014
Pages 105-111