Hidden diversity of Hepatozoon parasites infecting lizards from Central Brazil revealed by integrative taxonomy

The Brazilian Cerrado harbors one of the world’s richest lizard assemblages, yet the diversity and host associations of Hepatozoon parasites in this biome remain poorly understood. Here, we investigated Hepatozoon infections in free-ranging lizards using an integrative taxonomic approach combining morphology, morphometry, molecular screening, phylogenetic inference, and population genetic analyses. A total of 302 blood samples from 19 lizard species collected across three Cerrado localities were examined using microscopy and 18 S rRNA sequencing. Hepatozoon DNA was detected in 8.9% of the samples, with a subset of infections confirmed microscopically. Integrative analyses supported the formal description of Hepatozoon semitaeniati n. sp., representing the first record of hemogregarine infection in Tropidurus semitaeniatus and the first description of a Hepatozoon species infecting this host. A second lineage was recognized as a putative undescribed species based on concordant morphological and molecular evidence. We also confirmed the occurrence of Hepatozoon ameivae in Ameiva ameiva from Central Brazil, extending its known geographic distribution and providing the most complete 18 S rRNA sequence currently available for the species. Haplotype network, Bayesian clustering, and analysis of molecular variance (AMOVA) revealed marked genetic structuring among Brazilian Hepatozoon lineages, indicating clear differentiation between reptile- and rodent-associated parasites. Collectively, these findings reveal previously unrecognized diversity of Hepatozoon infecting Neotropical lizards and demonstrate the value of integrative taxonomy for resolving species boundaries, host associations, and evolutionary patterns of reptile-associated Hepatozoon .