Hoplias malabaricus comprises seven karyomorphs (A-G) and evolutionary units have been described in some of them. In this study, the karyotypic composition and genomic organization of individual H. malabaricus from Central Amazon are described and to verify whether they can be classified according to known karyomorphs. Individuals from the Ducke Reserve have 2n = 42 chromosomes, similar to karyomorph A. Individuals from Catalão Lake and Marchantaria Island exhibit 2n = 40 chromosomes, similar to karyomorph C. Regarding the constitutive heterochromatin, individuals from all locations present centromeric/pericentromeric blocks, in addition to some bitelomeric and interstitial markings. The number of chromosomes with nucleolar organizer region, 5S rDNA and 18S rDNA sites varied among the different locations. The Rex 3 element has a compartmentalized distribution at the terminal and centromeric regions of most chromosomes, with subtle differences among populations. Fluorescence in situ hybridization performed with a telomeric probe allowed the detection of these regions only at the terminal ends of the chromosomes. Thus, only the chromosomal macrostructure (karyomorphs A-G) is not sufficient to establish evolutionary units within the H. malabaricus group, considering differences in the genome organization that are found among their populations. Such differences in the genomic organization could be mainly caused by the sedentary habits of this species.
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