A modified SOS-Chromotest procedure to test for genotoxicity and cytotoxicity in sediments directly without extraction.
A modified SOS-Chromotest bioassay using a chromogenic pad (pad procedure) was developed to test for genotoxicity in sediments directly without extraction. This test is based on the de novo synthesis of beta-galactosidase enzyme by a genetically-engineered E. coli strain PQ37. In the bioassay, an exponential growth phase antibiotic-containing culture of the test bacterium is introduced into a series of tubes with the first tube containing 0.1 gram of sediment. Serial dilutions are then made and the tubes of sediment plus bacterial culture are incubated at 37 degrees C for four hours, followed by placing a drop of each mixture on a chromogenic pad and additional incubation for 20 hours at 37 degrees C. The solid particulates are then washed off with tap water and positive (genotoxic) activity is noted by the presence of a distinctive blue colour on the pad. The SOS-Chromotest pad procedure may be best used as a relative measure of genotoxicity by comparing results to a reference sample. In addition it can also determine sediment cytotoxicity by comparing samples spiked with a genotoxic standard (i.e., 4-nitroquinoline-N-oxide). Preliminary results suggest that this new bioassay is highly sensitive, consistent and discriminating.
D015112:4-Nitroquinoline-1-oxide / Q000737:chemistry; D001681:Biological Assay / Q000191:economics / Q000379:methods; D003124:Colorimetry; D003362:Cost-Benefit Analysis; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D005618:Fresh Water / Q000032:analysis / Q000737:chemistry; D005818:Genetic Engineering; D009152:Mutagenicity Tests / Q000191:economics; D009153:Mutagens / Q000032:analysis / Q000633:toxicity*; D013014:SOS Response, Genetics; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000633:toxicity*; D001616:beta-Galactosidase / Q000378:metabolism
D009153:Mutagens; D014874:Water Pollutants, Chemical; D015112:4-Nitroquinoline-1-oxide; D001616:beta-Galactosidase