Soil salinity and the salinity of trophic resources may alter the osmoregulatory processes of arthropod, challenging the smooth regulation of body water, and, ultimately, survival. The intra and extracellular build-up of osmolytes represent a common strategy to attenuate acute hyperosmotic stress in several arthropod species. In the present study, we aimed to determine the impact of substrate and trophic resource salinities on salt tolerance in the female wolf spider, Arctosa fulvolineata, which is considered a specialist salt marsh species. We evaluated adult female survival and egg laying, and quantified the osmo-induced accumulation of compatible solutes (GC-MS). Three concentrations of substrate salinity were tested (0‰, 35‰ and 70‰) under three trophic conditions (starved spiders, spiders fed with salt prey [intertidal amphipods] and spiders fed with unsalted prey [freshwater amphipods]). We found no support for diet preferences in female A. fulvolineata, which exhibited similar predation rates on freshwater and marine amphipods. Survival and egg-laying were significantly impaired when female A. fulvolineata were exposed to hypersaline conditions for 12 days. Our results showed an increase in the level of several compatible solutes when spiders were exposed for 12 days to saline conditions. For instance, α-alanine, β-alanine, arginine, asparagine, aspartate, homoserine, glutamine, glycine, proline and serine levels were 4-10 times higher under hypersaline conditions. The osmo-induced accumulation of amino acids may increase the osmolality of body fluids, thus enhancing the smooth regulation of body fluids and survival ability of wolf spider under extreme saline conditions.