Paternity misidentification is harmful due to the reduction in annual genetic earnings of the population and because it endangers an efficient genetic improvement program. The objectives of the present study was to evaluate nine microsatellites in Paternity Testing and to investigate misidentification paternity frequency in families of Gyr breed bovines population. In the present experiment blood samples from forty Gir breed families ( bull / cow / calf ), registered pure breed in the Zebu Breeders Brazilian Association (ABCZ) were used. The most part of the microsatellites used in this work were recommended by the International Society of Animal Genetics (ISAG). The genomic DNA extraction was performed from whole blood samples. The microsatellites TGLA122, TGLA126, BM1824, BMS2533, SPS115, ETH3, ETH10, ETH225 and POTCHA were amplified by PCR. The amplification products were separated by electrophoresis in denaturing polyacrylamide gel. from the obtained data, allele frequencies, Gene Diversity, Polymorphism Informative Content and Probability of Exclusion for each microsatellite marker were calculated. the genotype frequencies, Heterozygosity, Combined Probability of Exclusion and Probability of Paternity have also been calculated in the considered families. The Combined Exclusion Probability for all microsatellites was around 0.9789. The Paternity Testing results showed misidentification in eleven of the 40 studied families, that means, 27.5 percent of the sample. The Paternity Probability ranged from 0.8691 to 0.9999, and the mean was 0.9512