ABSTRACT Fatigue testing at ultrasonic frequency is a powerful method for rapid generation of high and very high cycle fatigue data. High accuracy of cyclic loading is the basis for reproducible test results, which can be obtained with closed-loop control of vibration amplitude and resonance frequency. Self-heating of test specimens can be suppressed by intermittent (pulse-pause) loading. In the present work, the high-accuracy ultrasonic fatigue testing equipment developed at the University of Natural Resources and Life Science, Vienna (BOKU) is described, and systematic investigations are presented that demonstrate the effects of cycling frequency, specimen size, and intermittent loading. Data obtained with additively manufactured (AMed) austenitic stainless steel 316L at 19 kHz are compared with electromagnetic resonation test results measured at 90 Hz. No influence of cycling frequency on fatigue lifetimes and fatigue limit was found when specimens of comparable size were used for tests at 19 kHz and 90 Hz. In contrast, the use of as-built specimens with smaller testing volume resulted in an increase in cyclic strength, demonstrating a size effect on fatigue properties. Polishing the surfaces of AMed specimens increased the fatigue limit by a factor of two. Furthermore, ultrasonic fatigue tests with intermittent loading and four different pulse lengths as well as continuous cycling were performed. The tests were conducted with wrought aluminum alloy 7075-T651 and with specimens containing small artificial surface defects. Similar lifetimes were measured in all testing series. Moreover, the same fatigue limit is found for all pulse lengths and for continuous cycling. This demonstrates that pulsed loading is a most suitable method to avoid specimen heating at ultrasonic frequency and that there is no influence on measured fatigue properties if ultrasonic loading is appropriately controlled. Measurement artifacts could be identified and avoided by measuring and recording all load amplitudes during ultrasonic fatigue tests.