Abstract: Chromosomal localization of sixteen bovine microsatellites in river buffalo has been assigned in this study, using polymerase chain reaction and buffalo-hamster somatic cell hybrids. These tested microsatellites were previously assigned to sheep chromosomes. This study also aimed to confirm that the chromosome band identity between river buffalo, cattle and sheep is a good indicator of genetic homology between these closely related species. The correlation coefficients between these tested microsatellites and other markers- representing syntenic groups and chromosomes in river buffalo- was calculated to assign these microsatellites to river buffalo chromosomes (BBU)). The results showed that BM719 is assigned to BBU5q, BM827 to BBU12, BM1818 to BBU2p, BM1824 to BBU1q, BM2113 to BBU2q, CSSM015 to BBU3p, CSSM034 to BBU4q, CSSM037 to BBU3q, CSSM043 to BBU1p, CSSM058 to BBU21, ILSTS011 to BBU15, ILSTS013 to BBU10, ILSTS019 to BBU5p, ILSTS029 to BBU6, ILSTS054 to BBU20 and ILSTS060 to BBU4p. The result also showed that 16 tested microsatellites are localized in river buffalo, cattle and sheep on the equivalent chromosomes, which have chromosome- band homology, in these closely related species belonging to Bovidae family, which has a high degree of chromosome conservation between its members and where the bi-armed autosomes are formed by centric fusions of acrocentric autosomes.