Alkylphenols On Marine Life Blood Lymphocytes Dna Damage Detected By Comet Assay

The comet assay, also known as the single cell gel electrophoresis (SCGE) assay, is a rapid and highly sensitive technique for measuring DNA breakage in individual cells. The technique has a wide range of applications due to its unique advantages such as quickness, high sensitivity, and small sample requirement. This study aims to establish SCGE technique detect the cellular DNA damage of shellfish blood hemolymph cells caused by alkylphenolics compounds, and further employ this technique in marine environmental risk assessment.The effects of nonylphenol,octylphenol,bisphenol A and butylphenol on DNA damage in shellfish hemolymph cells are studied at such exposure concentrations as 0,0.025,0.10,0.25,1.00mg/L. After 24hs and 48hs of exposure, the SCGE tests are carryed on to detect the cellular DNA damage.Firstly, the effect of nonylphenol on DNA damage in hemolymph cells of scallop was detected through in vivo and in vitro tests. Secondly the effects of nonylphenol on DNA damage in hemolymph cells of oyster and clam are studied. The results show that with the increase of NP concentration, comet tail increases obviously both in vivo and vitro assay. The higher the NP concentration is, the more serious the DNA damage is in shellfish hemolymph cells. There is a distinct dose-effect correlation between NP concentrations and the DNA damage levels in shellfish hemolymph cells. 0.025 mg/L is primarily defined as the genetic damage risk concentration of NP to shellfish. When the NP concentration exceeds 0.025 mg/L, the stability of DNA strands in shellfish will be affected and there will be a risk of carcinogenesis, mutagenesis and teratogenesis. Human health will also be affected due to seafood consumption. On the other hand, the DNA damage level and migration length show some difference among shellfish, oyster and clam exposed to the same nonylphenol concentration. This is because different shellfish species have different immunity to NP. But the mechanism for specific immunity difference to exogenous chemicals remains unknown both in China and overseas countries and further studies are required in the future.