The organization and mapping of multigene families can produce useful genetic markers, and its use may elucidate the mechanisms of karyotype variation and genomic organization in different groups of eukaryotes. To date, few species of Coleoptera have been analyzed using FISH for the location of multigene families. The purpose of this study was to use high-resolution chromosome mapping to establish the genomic organization of the 18S rDNA, 5S rDNA and histone H3 gene families in Lagria villosa. FISH was performed using 18S rDNA, 5S rDNA and histone H3 probes prepared via PCR labeling. Fiber-FISH for 18S and 5S rDNA indicated that both ribosomal elements are colocalized in the short arm of chromosome 4. Additionally, FISH, using the histone H3 probe, revealed that this sequence is found in only one autosomal pair and did not colocalize with rDNA. Fiber-FISH with 5S and 18S probes, used to improve the mapping resolution of these regions, showed that both genes are closely interspersed with varying amounts of both DNA classes.