We provide a karyological study of 12 species of the genus Pnigalio in an attempt to evaluate the taxonomic utility of karyotypes at the species level. For all species of Pnigalio examined the number of chromosome was 2n=12. Karyotype formulae presented mainly metacentric and submetacentric chromosomes, although a pair of acrocentrics or subtelocentrics, shorter than biarmed chromosomes, was present in some species. The analysis of karyotypes of Pnigalio showed frequent but not general interspecific variability of the chromosome traits. Although most of the studied species revealed concordance between morphological and karyological characters (centromeric index and relative length), two other categories have been identified: morphologically distinct species without reciprocal differences in karyotype structure, and morphologically similar species that strongly differ in chromosomal characters.