Simple SummaryPhysical fences are not always possible, thus automated technology called “virtual fencing” provides a potential solution. Virtual fencing uses Global Positioning System (GPS) technology and animals wear collar devices. As animals approach the virtual fence line, the collar emits an audio tone; if the animals walk further forward, they receive an electrical stimulus. If the animal turns around after the audio tone, they receive no electrical stimulus. However, no studies to date have looked at how animals respond when virtual fences have moved to different paddock locations. Virtual boundaries were set up to restrict six beef cattle wearing collars to different paddock areas. Within a few days, the animals were able to avoid the electrical stimulus by learning to turn away from the fence when they heard the audio tone. Over several weeks, the virtual fence was moved to three different locations within the paddock, and the animals rapidly learned it had moved, turning away at the audio tone the majority of the time. This shows that animals can learn the different collar signals and avoid moving virtual boundaries via the audio tone. The application of virtual fencing to farms enables improved animal management and animal exclusion from environmentally sensitive areas.AbstractGlobal Positioning System (GPS)-based virtual fences offer the potential to improve the management of grazing animals. Prototype collar devices utilising patented virtual fencing algorithms were placed on six Angus heifers in a 6.15 hectare paddock. After a “no fence” period, sequential, shifting virtual fences restricted the animals to 40%, 60%, and 80% of the paddock area widthways and 50% lengthways across 22 days. Audio cues signaled the virtual boundary, and were paired with electrical stimuli if the animals continued forward into the boundary. Within approximately 48 h, the cattle learned the 40% fence and were henceforth restricted to the subsequent inclusion zones a minimum of 96.70% (±standard error 0.01%) of the time. Over time, the animals increasingly stayed within the inclusion zones using audio cues alone, and on average, approached the new fence within 4.25 h. The animals were thus attentive to the audio cue, not the fence location. The time spent standing and lying and the number of steps were similar between inclusion zones (all p ≥ 0.42). More lying bouts occurred at the 80% and lengthways inclusion zones relative to “no fence” (p = 0.04). Further research should test different cattle groups in variable paddock settings and measure physiological welfare responses to the virtual fencing stimuli.