Background and Aim: Studies on avian influenza virus (AIV) in Libya are few and limited. This study aimed to determine the presence of AIV in live bird markets (LBMs) in Tripoli and determine the risk factors associated with AIV spread. Materials and Methods: In total, 269 cloacal swabs were randomly collected from different bird species in 9 LBMs located in Tripoli and its surrounding regions. The target species were ducks, geese, local chickens, Australian chickens, Brahma chickens, turkeys, pigeons, quails, peacock broiler chicks, and pet birds. Total RNA was extracted from the swab samples and used for real-time polymerase chain reaction to detect AIV type A. Results: Of the 269 samples, 28 (10.41% of total samples) were positive for AIV type A. The LBMs with positive samples were Souq Aljumaa, Souq Alkhamees, Souq Althulatha, and Souq Tajoura. The highest percentage (35.71%) of AIV was recorded in Souq Aljumaa. Positive results for AIV type A were obtained primarily in three species of birds: Ducks (14/65; highest percentage: 21.5%), local chickens (12/98; 12.24%), and geese (2/28; 7.14%). Furthermore, the following three risk factors associated with the spread of AIV type A were identified: Time spent by breeders/vendors at the market (odds ratio [OR] = 11.181; 95% confidence interval [CI] = 3.827-32.669), methods used for disposing dead birds (OR = 2.356; 95% Ci = 1.005-5.521), and last visited LBM (OR = 0.740; 95% CI = 0.580-0.944). Restricting the movement of poultry vendors from one market to another may protect against AIV spread. Conclusion: The findings of this study indicate the high risk of AIV spread in LBMs and highlight the need for continuous surveillance of LBMs across the country. Keywords: avian influenza, live birds market, risk factors, Tripoli.