Microsatellite markers previously developed for Pitcairnia albiflos Herb. and Pitcairnia geysksii L.B.Sm. were used in cross-amplification tests of five other Bromeliaceae species. Ten (76.9%) out of the 13 evaluated pair of primers had positive results for some of the species tested. Leaf samples were collected from individuals of these species in natural populations occurring in fragments of the Atlantic Forests, in Burarama, Cachoeiro de Itapemirim, ES. DNA of plant samples were extracted and purified using the cetyltrimethylammonium bromide (CTAB) extraction method, as described by Doyle & Doyle (1990).In order to improve the PCR result, the optimal annealing temperatures of each pair of primers to be tested were determined (between 48 and 56 °C). In all cases, microsatellite loci were amplified in a 15 µL volume containing 0.4 µM of each primer, 1 U Taq DNA polymerase, 0.1 mM of each dNTP, 1 × MgCl 2 -free reaction buffer (10 mM Tris-HCl pH 8.3 and 50 mM KCl), 2 mM MgCl 2 and 30 ng of template DNA.Amplifications were performed using a Techne TC-412 thermal cycler under the following conditions: 5 minutes denaturation at 94 °C followed by 30 cycles of 1 minute of initial denaturation at 94 °C, 1 minute of annealing temperature at 54 °C and 1 minute of extension at 72 °C, and elongation at 72 °C for 7 minutes.Amplified fragments were separated by electrophoresis on 2.5% agarose gel containing 0.02 ug/mL ethidium bromide, 1x TBE buffer (0.89 M Tris-HCl pH 8.3, 0.89 M boric acid and 0.02 M EDTA), at 110 volts for approximately three hours. Afterwards, the gels were photo graphed under UV light, using the gel documentation system Biolocus L PIX (Loccus Biotecnologia ® ). Gel electrophoresis was used to assess the number and size of amplified fragments, as well as polymorphism detection.Analyses of genetic variability at the microsatellite loci were done using the genotypes obtained for all five species of bromeliads evaluated in this study. The number of alleles per locus and the observed and expected heterozygosities under Hardy-Weinberg equilibrium were estimated. These analyses and the test for deviation from Hardy-Weinberg expectations were performed with Genes program (Cruz 2008).