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Seventeen trilobite species belonging to 16 genera and 13 families are systematically documented from the Houping Formation (upper Dapingian-lower Darriwilian, Middle Ordovician) at the Datangkou and Dacao sections, Chengkou, northern Chongqing. Of these, a new species, Mioptychopyge chengkouensis sp. nov., is established and described. New morphological information is provided for previously known species on the basis of new specimens. Two trilobite associations based on faunal composition and dominant forms are distinguished from the Houping Formation, in ascending order, the Agerina Association (from the lower and middle members) and the Nileus-Illaenus Association (from the upper member). The Agerina Association lived in an inner shelf margin environment with carbonate substrate above storm wave base (<70 m deep), whereas the Nileus-Illaenus Association developed in a shallow outer shelf environment below storm wave base (>70 m deep), indicating a rise of relative sea level corresponding to the early Darriwilian transgression. The cluster and nonmetric multidimensional scaling analyses of the trilobites in the formation show that the fauna had a strong endemicity to South China and a close biogeographic connection with east Peri-Gondwana, especially Tarim, Annamia and central Iran. [ABSTRACT FROM AUTHOR]