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Analysis of the toxicity of diflubenzuron and p-chloroaniline in biochemical indicators of non-target aquatic organims

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Author(s):
Darlene Denise Dantzger
Total Authors: 1
Document type: Master's Dissertation
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
Defense date:
Examining board members:
Hiroshi Aoyama; Vera Lucia Scherholz Salgado de Castro; Claudio Martin Jonsson
Advisor: Hiroshi Aoyama
Abstract

The use of agricultural products has been the main way to combat parasites in aquaculture, and the diflubenzuron (DFB) is the most used for that. This compound inhibits the chitin synthesis of the parasites exoskeleton, and has low toxicity to fishes. However, in the aquatic environment, the DFB can be toxic to non-target species and, when degraded, generate p-chloroaniline (PCA), a metabolite potentially carcinogenic and mutagenic to humans. According to requirement for more information about the toxicity of these compounds on aquatic organisms, the purpose of this study was to analyze the enzymatic activity of acid and alkaline phosphatases, catalase and superoxide dismutase for microalgae Pseudokirchneriella subcapitata, microcrustacean Daphnia similis and fish Oreochromis niloticus based on 50% effective concentration (EC50) for in vivo tests, and 50% inhibiting concentration (IC50) for in vitro tests, for the DFB and its metabolite PCA. The DFB EC50 was 0.00096 mg.L-1, 0.009 mg.L-1 and > 100 mg L-1 for algae, daphnia and fish in acute test. The PCA EC50 for daphnia and fish was 0.27 mg.L-1 and 24 mg.L-1, respectively. In turn, the algae had no change in their growth during exposure to PCA. In biochemical in vivo assays the DFB and PCA caused alterations in the activities of acid and alkaline phosphatases, catalase and superoxide dismutase for test organisms. Thus, the enzymes studied were sensitive to the exposure of these compounds in vivo studies and may be used as biomarkers of water pollution resources by DFB and PCA. On the in vitro test, the enzymes were incubated with the compounds DFB and PCA at different concentrations, only the FAT and CAT of fish liver had decreased activity with exposure to different concentrations of PCA, therefore, were used for determination of kinetic constants Km, Ki and Vmax, and IC50. The FAT had a competitive inhibition (Ki 8,2 mM - Km 0,21 mM - Vmax 0.01211 and IC50 3.2 mM), the CAT had a non-competitive inhibition (Ki 0,030 mM - Km 56,3 - Vmax 0.001198 and IC50 0.20 mM) when the concentration of PCA was increased. The in vitro test was important because has clarified the mechanism of action of PCA on FAT and CAT fish enzymes. The results of this study showed that both compounds were toxic to test organisms and caused significant changes in biomarkers evaluated. Therefore, the use of DFB must be done carefully (AU)

FAPESP's process: 11/04788-6 - Comparative toxicity of diflubenzuron and p-chloroaniline in phosphatases and antioxidant enzymes of non-target organisms
Grantee:Darlene Denise Dantzger
Support Opportunities: Scholarships in Brazil - Master