In order to characterise the telomeric repeats of the pufferfish Tetraodon fluviatilis, fluorescent in situ hybridization (FISH) was carried out on metaphase chromosomes using a PCR generated probe (TTAGGG)(n). Distinct signals have been observed on the telomeres of all the chromosomes and hybridization signals appears of uniform size and intensity. Moreover FISH experiment does not evidence any interstitial non-telomeric signal. As non-telomeric FISH signal has been used as a marker for karyotype evolution and for determining the evolutionary status of fish species, it could be supposed that the absence of non telomeric signals makes T. fluviatilis (Tetraodontiformes) an evolutionary ancient species in respect to Perciformes.