Six species of Eutardigrada are recorded from Portugal; four of them, Macrobiotus crenulatus Richters, 1904, Hypsibius seychellensis Pilato, Binda & Lisi, 2006, Diphascon (Diphascon) pingue (Marcus, 1936) and D. (Diphascon) patanei Binda & Pilato, 1971 are recorded for the first time in Portugal. Two species, Minibiotus orthofasciatus sp. nov. and Bertolanius (new name of Amphibolus) portucalensis sp. nov. are new to science. Minibiotus orthofasciatus sp. nov. is one of the species of the genus with three macroplacoids, microplacoid and cuticular pores forming transverse bands. The new species differs from all existing species by one or more of the following characters: distribution of the pores, shape of the pores, absence of dots on the legs, level of insertion of the stylet supports on the buccal tube. To the new species is attributed an unembryonated egg similar to those of Minibiotus intermedius (Plate, 1888), M. poricinctus Claxton, 1998, M. floriparus Claxton, 1998, and M. weglarskae Michalczyk, Kaczmarek & Claxton, 2005 but different from them in some details. Bertolanius portucalensis sp. nov. is very similar to the other species of the genus, but it differs from them in having very small cuticular tubercles. From some of them it differs by characters of the buccopharyngeal apparatus and/or of the eggs. This is the first record of the genus and of the Eohypsibiidae family in the Iberian Peninsula.