Background: Yemen recently experienced the largest cholera outbreak in the world, which started at the end of 2016. Cholera causes an estimated 1.3–4.3 million illnesses and 30,000–140,000 deaths worldwide each year, Cholera is caused by toxigenic Vibrio cholera serogroup O1 or O139 and is transmitted through fecal oral route. Infection is usually associated with drinking contaminated water and poor hygiene and sanitation (World Health Organization, 2019).   Objective: This study aimed to identify the risk factors for cholera transmission in the city of Sana' a, Yemen. Methods: We conducted an unmatched case control study. Cases were identified from the health unit records of Sana'a University and 230 participants were interviewed on socio-economic, household hygiene, food, and water exposure practices using a semi-structured questionnaire. Data were analyzed using SPSS version 22, and statistical significance was set at P < 0.05. Results: The 230 participants included 45 cholera cases (19.5 %) and 185 controls (80.5 %). Ages ranged from 15 to 65 years, with a mean of (20.0±47.0) years. According to the WHO, the cholera attack rate was 0.16 %, with no fatalities. Report 2017. Most participants 206 (89.5 %) had heard of cholera. Poor hygienic practices 199 (85.5.2 %) and contaminated water sources 159 (69.1 %) were the main reported transmission routes for cholera. Good hygienic practices 211 (91.7 %) were the main preventive methods for cholera in both cases, 45 (84.6 %)] and controls, 185 (89.0 %)]. The risk factors for cholera in the study population analysis showed Most participants 186 (80.8 %) practiced good food preservation methods. Moreover, 108 (46.9 %) ate outside their homes, such as in restaurants. Those with cholera were 11.2 times more likely to practice poor food preservation methods than those without the disease (p < 0.0001). Conclusions: Cholera transmission risk factors in Sana'a City were related to water and sanitation hygiene, and not washing khat before chewing is a specific risk factor for cholera; therefore, increasing awareness of the population on the importance of water chlorination and washing fruits, khat, and vegetables through a health education campaign is strongly recommended.