Background: West Nile Virus (WNV) is a zoonotic virus posing significant public health challenges globally. Misinformation about WNV on digital platforms like YouTube can impede public health efforts . Objective: This study evaluates the quality of English-language YouTube content on WNV using DISCERN and Global Quality Scale (GQS) tools to assess reliability and educational value. Methods: A YouTube search was conducted on September 21, 2024, using the keyword ”West Nile virus.” After applying inclusion and exclusion criteria, 79 videos were analyzed. Video characteristics such as source, type, and duration were assessed. DISCERN and GQS were used to evaluate quality and reliability and statistical analyses performed to identify factors influencing these scores. Results: The average DISCERN score of videos was 28, indicating low quality, with 64.5% of videos scoring 1 or 2 on the GQS. Videos uploaded by healthcare professionals or academic institutions had significantly higher GQS scores. Educational videos scored higher on both DISCERN and GQS metrics compared to testimonial or other videos. Video duration positively correlated with quality scores, while the presence of supporting visuals had no significant impact. Conclusion: Most YouTube content on WNV lacks reliability and quality, emphasizing the need for increased contributions from healthcare professionals and academic institutions. Producing professional, informative videos and encouraging viewers to rely on verified sources are critical for combating misinformation and improving public health communication.