Objectives: To investigate the knowledge and awareness of ultrasound practitioners’ concerning ultrasound artefacts in evaluating the hepatobiliary system. Methods: This electronic questionnaire-based comparative study involved the ultrasound practitioners’ who work in the radiology departments in Almadinah Almunawwarah governmental hospitals during the period from 1 November 2020 to 30 April 2021. Spearman’s rho correlation test was used to correlate between knowledge and job, academic qualification, and years of experience. A T-test and cross tabulation test were done to compare the knowledge about artefacts among radiologists and radiologic technologists. Results: This study involved 94 participants distributed as 22 (23.4%) radiologists and 72 (76.6%) radiologic technologists. The results shows that 85%, 71%, 73%, 69%, 54% and 53% of the participants assigned the acoustic shadowing, acoustic enhancement, ring down, side lobe, reverberation and mirror artefacts, as artefacts respectively. However, 68%, 53%, 19%, 19%, 18%, and 40% of the participants gave correct final diagnosis of acoustic shadowing, acoustic enhancement, ring down, side lobes, reverberation, and mirror artifacts, respectively. Spearman’s rho correlation test shows significant correlation between participants with more than three years experience and knowledge related mirror artefacts (r=0.328, p=0.001). It shows significant correlation between radiologists with knowledge related mirror artefacts (r=0.367, p<0.001). A significant correlation was found between highly qualified participants and knowledge related mirror artefacts (r=0.336, p=0.001) and side lobe artefacts (r=0.237, p=0.008). Conclusion: The questionnaire-based comparative study of knowledge about artefacts of hepatobiliary ultrasound imaging reveals a high level of Ultrasound practitioners’ knowledge in differentiating artefacts from pathology with a high level of knowledge in identifying hepatobiliary acoustic shadowing and acoustic enhancement artefacts. However, insufficient knowledge was noted in identifying mirror, side lobe, reverberation and ring down artefacts. A direct link was found between academic qualification, years of experience and practioners’ knowledge among. doi: https://doi.org/10.12669/pjms.38.6.5084 How to cite this:Alsaedi HI, Krsoom AM, Alshoabi SA, Alsharif WM. Investigation Study of Ultrasound Practitioners’ Awareness about Artefacts of Hepatobiliary Imaging in Almadinah Almunawwarah. Pak J Med Sci. 2022;38(6):1526-1533.  doi: https://doi.org/10.12669/pjms.38.6.5084 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.