Brucellosis, an important bacterial zoonosis caused by Brucella species, has drawn increased attention around the world. As an intracellular pathogen, the ability of Brucella to deal with stress within the host cell is closely related to its virulence. The survival pressure on Brucella within a phagosome is considered similar to that during the stationary phase. Here, label-free proteomics approach was used to study the adaptive response of Brucella abortus (B. abortus) in the stationary stage. 182 down-regulated and 140 up-regulated proteins were found in the stationary-phase B. abortus. B. abortus adapted to adverse environmental changes by regulating virulence, reproduction, transcription, translation, stress response, and energy production. In addition, both logarithmic and stationary-phase B. abortus were treated with short-term starvation. The logarithmic-phase B. abortus restricted cell reproduction and energy utilization in response to nutritional stress. Additionally, the expression levels of some virulence-related proteins were identified as being significantly regulated during the transition from logarithmic to stationary phase or under starvation treatment, such as Type IV secretion system protein (T4SS), VjbR, and integration host factor (IHF). Altogether, we outlined adaptive mechanisms that B. abortus could employ during the growth and compared the differences between logarithmic and stationary-phase B. abortus in response to starvation.