The Tuotuo River is a tributary to the Yangtze River, and ecologically important because it supports diverse habitat including many endemic species of the Qinghai-Tibet Plateau. The Tuotuo River basin is especially unique and includes many different and fragile habitats, but bird and mammal diversity remain unclear. From September 2017 to December 2018, 60 infrared cameras were placed evenly in a grid (60 1-km 2 grids) across three sample plots, Bande Mountains, Daerzong Mountains and the mouth of Tongtian River. A total of 4,288 camera-days, and 2,353 independent photos were obtained. Based on these data, we identified a total of 38 species. We identified 16 species of mammals which could be divided into 5 orders, 10 families, 16 genera and 22 species of birds which could be divided into 8 orders, 15 families, 19 genera. Among 38 species, 26 are classified as falling under either first or second class animal protections in China or have important economic or research value. Four species that belong under Class-I National Key Protected Species include the snow leopard (Panthera uncia), white-lipped deer (Gervus albirostris), kiang (Equus kiang) and lammergeier (Gypaetus barbatus); 11 species are classified as Class-II National Key Protected Species and 11 species which are beneficial or have important economic value or research value. The saker falcon (Falco cherrug), a IUCN red list threatened and endangered species was detected, and two other vulnerable species and five species as near threatened were identified in images. The five most detected mammals in the Tuotuo River basin were blue sheep (Pseudois nayaur, RAI = 11.17), plateau pika (Ochotona curzoniae, RAI = 6.48), woolly hare (Lepus oiostolus, RAI = 4.99), Tibetan fox (Vulpes ferrilata, RAI = 3.36) and Tibetan gazelle (Procapra picticaudata, RAI = 2.87). The five most abundant avian species were red-billed chough (Pyrrhocorax pyrrhocorax, RAI = 3.08), black redstart (Phoenicurus ochruros, RAI = 1.26), rufous-necked snowfinch (Pyrgilauda ruficollis, RAI = 1.26), upland buzzard (Buteo hemilasius, RAI = 0.82) and white-rumped snowfinch (Pyrgilauda taczanowskii, RAI = 0.61). Meanwhile, livestock and human activities were also commonly recorded by infrared cameras (RAI = 8.72). The results provide an important basis for further investigating the biodiversity of the Tuotuo River basin of the Yangtze River and formulating conservation and management measures in the future.