A basic assay that detects genotoxic DNA damage disrupting DNA replication and repair mechanisms is the sister chromatid exchange test. The frequency of sister chromatid exchanges was analyzed in chromosomes of the following hen breeds: Greenleg Partridge and Polbar. Chromosome preparations were obtained from our in vitro culture of peripheral blood lymphocytes stained using the fluorescence plus Giemsa (FPG) technique. The sister chromatid exchange (SCE)/cell mean of the hens under analysis was 7.83 ± 1.76 (7.22 ± 1.70 in the Greenleg Partridge and 8.43 ± 1.61 in the Polbar population). Statistically significant differences were identified between the hen breeds. A higher mean number of SCE/cell was observed in the group of hens producing fewer eggs (8.55 ± 1.51) compared with the group with a better egg yield (7.10 ± 1.65). The differences were statistically significant. Additionally, SCE frequency in the first, second, and third chromosome was analyzed in detail. The highest number of SCE was observed in the first and the lowest in the third chromosome. The SCE distribution in the particular regions of the analyzed chromosomes was also studied. The most numerous exchanges were observed in the proximal region, followed by the interstitial and distal areas.