The Microtox test uses the marine luminescent bacterium, Photobacterium phosphoreum. To provide the organism with osmotic protection, sodium chloride (2.0%) is usually added to freshwater samples before analysis. This procedure is shown to cause a dramatic lowering of measured toxicity where ionic compounds are the major toxicants present. The reason for this is the consequent variation in ionic strength upon addition of NaCl rather than formation of less-toxic chloro complexes. Data are given for the toxicants Zn, Cd, Cu, NH4+, and phenol.It was found that measurements on aqueous samples could be made without adjustment provided that the salinity was in the range of 1.0-7.0%. Freshwaters can also be analyzed without adjustment of ionic strength by adding sucrose (20.4%) to provide the necessary osmotic protection.It is therefore recommended that the ionic strength of aqueous samples should not be altered as part of the test and that receiving waters should be used as the diluent wherever possible.