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Erod measurement using post mitochondrial supernatant (pms) in roach (Rutilus rutilus L.), a possible biomonitor for
D B O'Hare1, P W Robotham, R Gill
1School of Environmental and Applied Sciences, Department of Biology, University of Derby, U.K.
Abstract:
The use of hepatic post-mitochondrial supernatant (pms) as a source of monooxygenase activity in roach following intra peritoneal injection of beta-naphthoflavone was investigated. Pms was found to be three times less active than microsomes although the level of induction was similar (9 fold). No effect of dicumarol on Ethoxyresorufin O-deethylase (EROD) measurement using pms was recorded. EROD induction in roach peaked 2-4 days post treatment with beta-naphthoflavone. Given the ubiquitous nature and pollution tolerance of roach, their hepatic pms may serve as a convenient biomonitor of PAH contamination in the freshwater environment. Field studies are now underway to investigate this further.
Insights
Hepatic post-mitochondrial supernatant (pms) from beta-naphthoflavone-treated roach shows potential as a biomonitor for polycyclic aromatic hydrocarbon (PAH) contamination. While less active than microsomes, roach pms offers a convenient method for environmental monitoring.
Area of Science:
- Environmental toxicology
- Biochemistry
- Aquatic ecotoxicology
Background:
- Cytochrome P450 enzymes, particularly monooxygenases, are crucial for metabolizing environmental contaminants.
- Beta-naphthoflavone is a known inducer of these enzymes.
- Roach (Rutilus rutilus) are widespread freshwater fish, often exposed to pollutants.
Purpose of the Study:
- To investigate the utility of hepatic post-mitochondrial supernatant (pms) from roach as a source of monooxygenase activity.
- To assess the potential of roach pms for biomonitoring polycyclic aromatic hydrocarbon (PAH) contamination.
Main Methods:
- Intraperitoneal injection of beta-naphthoflavone into roach.
- Preparation of hepatic post-mitochondrial supernatant (pms) and microsomes.
- Assay of Ethoxyresorufin O-deethylase (EROD) activity.
- Evaluation of dicumarol's effect on EROD measurements.
Main Results:
- Hepatic pms exhibited lower monooxygenase activity (threefold less) compared to microsomes, despite similar induction levels (ninefold).
- Dicumarol did not affect EROD measurements using pms.
- EROD induction in roach peaked 2-4 days after beta-naphthoflavone administration.
Conclusions:
- Roach hepatic pms can be used to measure EROD activity, reflecting monooxygenase induction.
- The sensitivity and timing of EROD induction in roach suggest their hepatic pms is a viable biomonitor for PAH pollution.
- Further field studies are recommended to validate roach pms as an environmental biomonitoring tool.