Titanium dioxide (TiO2) nanoparticles have a high surface-to-mass ratio and rapidly aggregate in water causing great difficulties for toxicity test exposed to aquatic organisms or other cell lines. This study uses a cell viability kit for routine toxicity test of TiO2 as well as other nanoparticles which accumulate in the aquatic environment. Tetrahymena immediately endocytoses TiO2 nanoparticles and stores them in food vacuoles until the particles undergo exocytosis as larger aggregates. However, during the process of endocytosis and exocytosis, TiO2 particles interfere with cell growth and consequently induce acute toxicity. It exerted high cell growth inhibition at 20 h incubation and induces significant cytotoxic effects. Surprisingly, the effect of TiO2 decreases at 40 h incubation, due to the recovery of cell growth and reduction of the cytotoxicity of the particles.

A 96-well plate culture methodology for the unicellular eukaryote Tetrahymena pyriformis, strain GL was used for the determination of toxicity and metabolism of catecholamines. Catecholamines exhibited moderate acute toxicity to Tetrahymena cells where dopamine and L-DOPA showed higher toxic potential at EC(10) (0.39 ppm and 0.63 ppm, respectively) and EC(20) (1.1 ppm and 1.0 ppm respectively) after 48 h exposure. All tested catecholamines were highly degradable in the PPY-medium due to the oxidizing environment during incubation. Also the catecholamines were naturally synthesized and released by Tetrahymena cells into the culture medium and increasingly accumulated with time where noradrenalin exhibited the highest degree of accumulation. However, the exogenous exposure of catecholamines to the cells caused the depletion of natural noradrenalin synthesis even with the addition of very low physiological concentration (0.12 ppm). Dopamine caused the higher effect on inhibiting noradrenalin synthesis. Treatment with a higher concentration (8.0 ppm) of dopamine in 96-well plates caused strong excitation of the cells and ascertained a new metabolite in vivo while the other representative catecholamines were not responsible for the production of this metabolite. This dopamine metabolite is relatively non-polar as compared to noradrenalin, adrenaline and dopamine and eluting later through the reverse phase C-18 column.

Two types dopamine receptor present in the cell membrane of vertebrates. But in this study D1 receptor was identified in the invertebrate ciliates protozoan, Tetrahymena thermophila by use of fluorescent ligands. D1 specific agonist SKF-38393 binds specifically to Tetrahymena. The specific binding of SKF-38393 was encountered by equimolar addition of D1 antagonist thus showed no binding of ligands. In addition, it was also proved that the D1 specific agonist did not cross bind with the D2 type receptor due to the equimolar addition of D2 selective antagonist spiperone. Interestingly this study also showed that the dopamine receptor present in the endoplasmic reticulum and endosomes of Tetrahymena as well as cell membrane which was revealed by laser scanning microscope. Therefore, this evidence supports the existence of a D1 receptor in the ciliate protozoan.

The present study has highlighted several future research directions. For ecotoxicological risk assessment of FTOH, their distribution and environmental fate should be determined. To understand the effect mechanism, more tests could be conducted to test whether apoptosis is caused. Finally, in order to standardize test procedure in a closed system, more compounds should be investigated in the closed system to clarify the sensitivity of the test procedures.