A puncture needle detection unit that detects an image of a puncture needle displayed in an ultrasound image, a motion extraction unit that extracts a linear motion at a point of the puncture needle based on a history of the image of the puncture needle in the ultrasound image, and a composite image generation unit that generates a composite image by generating a locus of the linear motion extracted by the motion extraction unit and superimposing the locus on the ultrasound image are included. With the configuration, an ultrasound observation apparatus, a method of operating an ultrasound observation apparatus, and an ultrasound observation apparatus operation program, which enable a user to grasp a position where the puncture needle has been moved a plurality of times in a subject, are provided.
