Detection of Metallothionein Protein Biomarkers (MTs) and Pinocytosis Activity in Gambusia Fish (Gambusia affinis) Exposed to Cadmium
2019
Moh. Awaludin Adam, Maftuch, Yuni Kilawati and Yenny Risjani
This study aims at detecting the metallothionein protein that binds cadmium in Gambusia fish using MT ELISA kit and Western Blotting testing. The continuation of the detection is used as a basis for analysing the macrophage and pinocytosis activity on Gambusia fish exposed to cadmium. The methods used to detect metallothionein protein are the MT ELISA kit and Western Blotting test. Meanwhile, to check the fish immunity, macrophage cells and pinocytosis activity were tested. The results indicated that the total proteins detected by the method of MT ELISA kits showed an increasing number with concentrations of exposure to Cd good at acute doses and sub-chronic higher doses (treatment A: 0.066 ± 0.019 ng/mL; B: 0.053 ± 0.022 ng/mL; C: 0,045 ± 0,014 ng/mL; D : 0,021 ± 0,012 ng/mL; E : 0,032 ± 0,019 ng/mL; and control k : 0,018 ± 0,018 ng/mL). Similarly, in the pinocytosis activity, the number of macrophage cells between treatment and control was significantly different (Cd ³ 3x106 treatment; Cd ³ 1x106 no treatment) and the number of pinocytosis activity was significantly different (Cd ³ 24 PA treatment; Cd ³ 10 no PA treatment). Increasing pollutants from cadmium exposure affect the fish health. Pollution causes changes in macrophage’s immune cells and phagocytosis Gambusia affinis, which can change the overall innate immunity system and fish health. The change of immune cells associated with the species and biological conditions is not present. Analysis of the number of immune cells revealed that Gambusia has decreased its macrophage pinocytosis activity, thereby indicating that phagocyte activity is a sensitive biomarker and a good bio-indicator for cadmium pollution.
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