The F1 hybrid offspring of the distant hybridization between Cichlasoma trimaculatum (male) and Heros managuense (female) were successfully obtained through artificial fertilization, and the chromosome specimen of kidney cells of the two parents and their offspring were prepared using a low colchicine concentration air drying method, Howell and Black's (1980)
silver–nucleolar organizer region (Ag–NOR) staining method and Sumner's (1972)
C-band staining method. The karyotypes, Ag–NORs and C-bands of C. trimaculatum, H. managuense and their offspring were observed. The results showed that the diploid numbers of the two parents and the F1 hybrid offspring were all 2n=48. For C. trimaculatum, the karyotype formula was 2n=2m+8sm+26st+12t, NF=58. Two Ag–NORs were seen on the tips of the short arm of chromosome st7, and the entire chromosomes of st9 and st13 and the centromeres of the other chromosomes had positive C-bands. For H. managuense, the karyotype formula was 2n=2m+6sm+28st+12t, NF=56. Two Ag–NORs were located on the tips of the short arm of chromosome sm2 and positive C-bands were found in the centromere of all chromosomes. In the F1 hybrid offspring, the chromosome formula was 2n=2m+8sm+30st+8t, NF=58. Two Ag–NORs were seen on the tips of the short arm of chromosome sm2; all chromosomes presented positive C-bands in the centromere. No heteromorphic sex chromosome was found. After comparison with the two parents, the F1 hybrid offspring was suggested to be the filial generation of the two hybrid parents rather than the gynogenesis of H. managuense induced by heterogeneous sperm of C. trimaculatum.