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Audio information plays a rather important role in the increasing digital content that is available today, resulting in a need for methodologies that automatically analyze such content: audio event recognition for home automations and surveillance systems, speech recognition, music information retrieval, multimodal analysis (e.g. audio-visual analysis of online videos for content-based recommendation), etc. This paper presents pyAudioAnalysis, an open-source Python library that provides a wide range of audio analysis procedures including: feature extraction, classification of audio signals, supervised and unsupervised segmentation and content visualization. pyAudioAnalysis is licensed under the Apache License and is available at GitHub (https://github.com/tyiannak/pyAudioAnalysis/). Here we present the theoretical background behind the wide range of the implemented methodologies, along with evaluation metrics for some of the methods. pyAudioAnalysis has been already used in several audio analysis research applications: smart-home functionalities through audio event detection, speech emotion recognition, depression classification based on audio-visual features, music segmentation, multimodal content-based movie recommendation and health applications (e.g. monitoring eating habits). The feedback provided from all these particular audio applications has led to practical enhancement of the library.