Objective To study the species and distribution of rodents and blood-sucking insects in Xisha Islands of China, so as to provide evidence for prevention and control of infectious diseases in the area. Methods Using mouse cages and light trap, we collected rodents and blood-sucking insects (mosquito and midge) in Yongxing and Shi Islands (two islands of China's Xisha Islands) during November to December in 2013 and February to March in 2014. The specimens were identified by morphologic characters and molecular markers. Results A total of 160 rodent individuals, belonging to 3 species, were captured: Rattus norvegicus, Rattus tanezumi and Sorex sp, with Rattus norvegicu (61.25%) being the dominant species. The 315 collected mosquito samples belonged to 4 genera and 5 species, with the dominant species being Armigeres subalbatus (52.38%). The Anopheles mosquito was identified as Anopheles subpictus cytotype B by ITS2 sequence. The 121 collected midges belonged to 3 subfamilies, 4 genera and 8 species, with Culicoides peliliouensis (61.98%) being the dominant species. Conclusion The species numbers of rodents, mosquitoes, and midges in Yongxing and Shi Islands are 3, 5, and 8, respectively. This is the first record of Anopheles mosquitoes and midges in the area, and Forcipomyia (Lepidohelea) chrysolopha and Forcipomyia (Schizoforcipomyia) borbonicawere are firstly recorded in China.