Approved Abstracts

Post mortem UGT activity in green turtles tissues: implications for environmental biomonitoring

Author(s): Dias VHV; Mattos JJ; Bastolla CLV; Rogério DW; Jorge JHB; Bainy ACD;
Presenter: Vera Helena Vidal Dias

The tissues of sea turtles, endangered species and or large vertebrates collected until 24 h after death attends legal, ethical, and logistic criteria for biomarker studies. However, how the post mortem period could affect the biomarker responses is not well understood. Uridine diphosphate glucuronosyltransferase (UGT) enzyme belongs to the phase II biotransformation systems. UGT is also considered as a biomarker of exposure because it protects the organism against toxic effects of contaminants. This study evaluated the post mortem UGT activity for up to 24 h in liver, kidney and small intestine of green turtles (Chelonia mydas). Samples were obtained from two juvenile female specimens of C. mydas (Cm A e Cm B), which were submitted to rehabilitation in TAMAR (Florianópolis, Southern Brazil) and euthanized due to the impossibility of returning into the environment. After sampling, all tissues were immediately frozen in liquid nitrogen and stored at -80ºC. Reactions were performed in triplicates by incubating microsomal samples, alamethicin, bovine serum albumin and Mg+2. UDPGA was added, and the microplate was transferred to a water bath at 30°C for 5 min. The reaction started by the addition of 4-MU. UGT activity was compared by one-way ANOVA followed by Dunnett's post hoc test. The level of significance was p< 0.05. In all tissues and specimens, 24 hours after death, UGT activities were smaller (p < 0,0001) than initial time. In the small intestine during all post mortem periods there was a decrease (p < 0.05) in UGT activity. However, some post mortem periods in liver and renal tissues showed an increase (p < 0.05) in UGT activity. Liver UGT activities of Cm A were higher at 2 h, 3 h and 18 h after death than initial time and 1 h after death of Cm B. Renal UGT activities of Cm A were higher at 12 h after death than initial time, and of Cm B at 1h and 2 h after death. The highest specific UGT activity was observed in kidney samples. The instability observed in the UGT activity during the post mortem interval could be related to the quantity of dead cells and cellular heterogeneity in sessions of the sample tissues. Furthermore, it is important to consider that these two specimens faced different treatments during rehabilitation for two or three months before the death. This work provides interesting information to the understanding of how UGT activity is affected after 24h of the post mortem period. This knowledge is useful for biomonitoring programs that use wild animals’ carcasses for biomarker analysis. We can conclude that it is very important to analyze tissues sampled as soon as possible after the death of the organism in order to get more reliable results. This study was supported by CNPQ Universal 425840/2016-6.

Keywords: biomonitoring; UGT activity ; Chelonia mydas




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