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Mice emit surprisingly complex ultrasonic vocalizations (USVs), often employed as a behavioural readout in various fields of biomedical science. Strong scientific efforts aim at understanding of the functions of mouse USVs, however most acoustic studies have very specialised aims and evaluate the behaviour only within short (often 5-10 minute) and not always biologically relevant experimental tests. Therefore, we conducted a pilot long-term study of the acoustic activity of socially established female mouse groups housed in enriched modular environment, using synchronised recording of audio and video data. With frame-by-frame video analysis we tested the effect of behavioural context on the time-frequency parameters of USVs and the occurrence of different call types. We found that female mice usually produce USVs during direct social interactions. Most of the call parameters showed a strong relation with context, however the largest changes during the transition from peaceful to aggressive interactions were found in the fundamental frequency (decrease), duration (increase), proportion of calling (increase) and the presence of nonlinear phenomena (increase). Since these changes in mice USVs (except for fundamental frequency) align with those obtained for audible calls of many mammals with increase in arousal, they may reflect some general biological rule that applies to both audible and ultrasonic signals. In the future, low-frequency, prolonged USVs with a high proportion of non-linear phenomena can potentially be used as non-invasive indicators of increased negative emotional expression in groups of female mice in home cages for a wide range of applied tasks.