SUMMARY.The livers of the marsupials Trichosurus vulpecula and Phascolomis mitchelli, the monotreme Echidna aculeata, the lizard Tiliqua rugosa and the frog Rana temporaria have been studied with regard to their content of arylsulphatases and related enzymes. Type I arylsulphatases were found only in the opossum (T. vulpecula) and the lizard, in very small amounts in the latter. The enzymes were generally similar to the type I arylsulphatases of the eutherian mammals although they differed in their considerably lower pH optima which were in the region of pH 6. Type II arylsulphatases were found in all the species studied. The enzymes had the electrophoretic properties of a sulphatase B from a eutherian mammal but they were not typical of this group as their hydrolyses of nitrophenyl sulphate were not greatly activated by Cl~ ions and they seemed to combine many of the properties of the sulphatases A and B of the eutheria. It is therefore suggested that the complexity of the arylsulphatases in the eutheria is a recent development in their evolution. No steroid sulphatase could be detected in opossum liver.Opossum liver could synthesise p-nitrophenyl sulphate only extremely slowly and no synthesis of androstenolone, oestrone or testosterone sulphates could be detected. Normal opossum urine contained no detectable ester sulphates. A dose of 1-naphthol was excreted by the opossum mainly in the form of urinary glucuronides and only traces of ester sulphates were formed.INTRODUCTION.