In recent years, bat-associated pathogens, such as 2019 novel coronavirus, have been ravaging the world, and ectoparasites of bats have received increasing attention. Penicillidia jenynsii is a member of the family Nycteribiidae which is a group of specialized ectoparasites of bats. In this study, the complete mitochondrial genome of P. jenynsii was sequenced for the first time and a comprehensive phylogenetic analysis of the superfamily Hippoboscoidea was conducted. The complete mitochondrial genome of P. jenynsii is 16 165 base pairs (bp) in size, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes and 1 control region. The phylogenetic analysis based on 13 PCGs of the superfamily Hippoboscoidea known from the NCBI supported the monophyly of the family Nycteribiidae, and the family Nycteribiidae was a sister group with the family Streblidae. This study not only provided molecular data for the identification of P. jenynsii, but also provided a reference for the phylogenetic analysis of the superfamily Hippoboscoidea.

The family Streblidae is a significant grouping of dipteran insects within the superfamily Hippoboscoidea, which parasitizes the body surface of bats. With the global spread of bat-related pathogens in recent years, Streblidae has gained increasing attention due to its potential for pathogen transmission. A sample of Brachytarsina amboinensis was sequenced on the B. amboinensis were obtained, compared with available Streblidae mitogenomes, and the phylogeny of Hippoboscoidea was reconstructed. The results indicate that the mitochondrial genome of B. amboinensis exhibits a relatively high degree of conservation, with an identical gene count, arrangement, and orientation as the ancestral insect's genome. Base composition analysis revealed a strong bias towards A and T in the base composition. Selection pressure analysis indicated strong purifying selection acting on cox1. Pairwise genetic distance analysis showed that cox1 evolved at a relatively slow rate. Regarding phylogenetic relationships, the constructed phylogenetic trees using Bayesian inference and Maximum Likelihood methods supported the monophyly of the Hippoboscoidea, Glossinidae, Hippoboscidae, and Nycteribiidae clades, with high nodal support values. Our research confirmed the paraphyly of the families Streblidae. In the familial relations between Nycteribiidae and Streblidae, New World Streblidae share a closer kinship with Nycteribiidae. This contrasts with prior findings which indicated that Old World Streblidae share a closer kinship with Nycteribiidae. This study not only enhances the molecular database for bat flies but also provides a valuable reference for the identification and phylogenetic analysis of Streblidae.

Species of the family Nycteribiidae are blood-sucking ectoparasites that parasitize bats. To further enrich the molecular data of species in the family Nycteribiidae, the complete mitochondrial genome of Nycteribia parvula was sequenced for the first time in this study. The complete mitochondrial genome of N. parvula is 16,060 base pairs (bp) in size, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, two ribosomal RNA genes, and a control region. The nucleotide contents of A, T, G, and C are respectively 40.86%, 42.19%, 6.51%, and 10.44%. The phylogenetic analysis based on 13 PCGs supports the monophyly of the family Nycteribiidae, and N. parvula is the closest relative to Phthiridium szechuanum.