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We conducted a comparative population genetic analysis of levels of genetic variation and its geographical structuring in three closely related species of grasshopper that co-occur in the Mediterranean Basin: Calliptamus italicus, C. barbarus and C. wattenwylianus. In the western part of their distributions 5 populations of C. italicus, 13 of C. barbarus and 10 of C. wattenwylianus were sampled. Bootstrap re-sampling of populations and microsatellite loci within each species indicated a lower level of genetic diversity and higher level of genetic differentiation in C. barbarus, which is less of an outbreak pest species than either of the other Calliptamus species studied. This may be due to lower effective sizes of non-outbreak populations and/or lower gene flow among them.