Gene expression of acetylcholinesterase in hybrid catfish (Clarias gariepinus X Clarias macrocephalus) exposed to chlorpyrifos and carbaryl.

After serving as a neurotransmitter, acetylcholine is hydrolyzed by acetylcholinesterase (AChE). Inhibition of AChE is considered to be a specific biomarker for exposure to organophosphate and carbamate insecticides. In this study, the AChE gene was isolated from hybrid catfish brain by RT-PCR methods using degenerate primers. The amplified fragment of 966 nucleotides generated by PCR was cloned, and sequence analysis showed 82% nucleotide identity with AChE of the Electrophorus electricus. Specific primers of the hybrid catfish AChE gene were then synthesized and used in the examination of AChE gene expression in brain tissue of hybrid catfish exposed to sublethal concentrations of chlorpyrifos (0.43, 4.3 and 43 2M) and carbaryl (1.19, 11.9 and 119 2M) for 24 hr. Real-time PCR was used to compare with the amplified 28S rRNA gene. AChE gene expression was significantly elevated 12.4 times in catfish exposed to 43 2M chlorpyrifos in comparison to the control group (p<0.05). Carbaryl did not produce any significant change. These results indicate that AChE is more sensitive to administration of chlorpyrifos than carbaryl. The induction of the AChE gene indicates the possibility of using this gene as biomarker for detecting effects of organophosphate insecticides in hybrid catfish.

Cytochrome P450 1A activity in liver and fixed wavelength fluorescence detection of polycyclic aromatic hydrocarbons in the bile of tongue-fish (Cynoglossus acrolepidotus, Bleeker) in relation to petroleum hydrocarbons in the eastern Gulf of Thailand.

This investigation was conducted in an area of oil spill along the east coast of Thailand to examine the relations among cytochrome P450 1A activity in liver and PAHs in the bile of the tonguefish and petroleum hydrocarbons in the sediments. PAH sediment concentrations in the reference and oil spill areas were 5.03 +/- 0.42 and 0.21 +/- 0.043 microg(-1) dry weight respectively Cytochrome activity in fish liver from oil spill area was 45.40 +/- 3.50 pmoles/ min/mg protein, almost threefold higher than that from the reference sites. Flourescense detection in bile metabolites at the oil spill area, 69.8 +/- 9.9 flourescense unit was significantly higher than that at the reference sites, 22.9 +/- 5.5 and 22.2 +/- 3.5 fluorescence unit. A strong correlation was found among cytochrome P450 1A activity in liver, PAH of bile metabolites and petroleum hydrocarbons. Both cytochrome and bile metabolites activity decreased seaward varying to the distance from the oil polluted area. We concluded that both detections in tonguefish can be regarded as a complementary biomarkers for the exposure of PAHs in tropical marine environments.