Although many physiological characteristics are very labile, it is proposed that some are stable enough to provide evidence concerning the routes by which terrestrial animals moved on to land. Osmotic tolerance and osmoregulatory ability are investigated first. Marine littoral invertebrates tolerate wide osmotic changes but osmoregulate little. Freshwater invertebrates osmoregulate well, but over a narrow range. Terrestrial prosobranchs of the family Pomatiasidae, which have wide tolerance, may have originated directly from the sea. The Cyclophoridae, which are intolerant, may have moved to land from fresh water. Isopods and talitrid amphipods may all have had direct marine origins with no freshwater phase. Secondly, terrestrial animals of marine origin may have blood with higher osmotic pressures than those passing through fresh water. In prosobranchs, isopods and amphipods, the evidence agrees with the conclusions suggested from character one. Thirdly, the production of hypo-osmotic urine in terrestrial animals may be evidence of freshwater origin. The evidence again supports the initial suggestions about prosobranchs and amphipods. It is concluded that because there is little selection pressure acting on these characters on land, they can be used as evidence of ancestry. Possible use of further characters is discussed.