In this paper we summarize for the first time the effects of agriculture expansion and intensification on animal diversity in the Pampas of Argentina and discuss research needs for biodiversity conservation in the area. The Pampas experienced little human intervention until the last decades of the 19th century. Agriculture expanded quickly during the 20th century, transforming grasslands into cropland and pasture lands and converting the landscape into a mosaic of natural fragments, agricultural fields, and linear habitats. In the 1980s, agriculture intensification and replacement of cattle grazingcropping systems by continuous cropping promoted a renewed homogenisation of the most productive areas. Birds and carnivores were more strongly affected than rodents and insects, but responses varied within groups: (a) the geographic ranges and/or abundances of many native species were reduced, including those of carnivores, herbivores, and specialist species (grassland-adapted birds and rodents, and probably specialized pollinators), sometimes leading to regional extinction (birds and large carnivores), (b) other native species were unaffected (birds) or benefited (bird, rodent and possibly generalist pollinator and crop-associated insect species), (c) novel species were introduced, thus increasing species richness of most groups (26% of non-rodent mammals, 11.1% of rodents, 6.2% of birds, 0.8% of pollinators). Much taxonomic and ecological work is still needed to understand Pampean animal biodiversity, to understand how agriculturization is affecting it, and to identify appropriate conservation actions. Networks of Important Bird Areas and Valuable Grassland Areas harbor a balanced representation of Pampean biodiversity and, if adequately protected, may provide valuable research sites, but complementary work should be carried out on agriculturized areas.