Cholera is an acute bacterial enteric disease characterized in its severe form by sudden onset, profuse painless watery stools, nausea, and vomiting. Aim of Study: to assess the knowledge, attitude and practice of female's employees about cholera at Sanaa university, Yemen. Subject and Method: descriptive cross sectional research design was carried out in the study. Sample: include 230 female's employees at Sanaa university Yemen. Tools: Interview questionnaire was used which include three tools have been used first tool which include two parts: part one includes sociodemographic characteristics, part two used to assess knowledge about cholera, second and third tools used to assess attitude and reported practice of females about cholera disease. Result: the study clear that age of female participant means ± SD was 32.85± 5.84, poor level of knowledge about cholera was observed among (75.5%), the majority of study sample had negative attitude toward cholera while only 30% of them had poor practice. Also, there is statistically significant difference between level of education of female's employee with their knowledge and practice. Conclusion: The study revealed that poor level of knowledge, negative attitude and poor practice about cholera among female's employees at Sanaa university in Yemen. Recommendations: The present study recommended implement awareness campaigns and educational programs in universities to estimate the level of knowledge toward cholera disease, also vaccination against cholera, and to warn from the hazards of cholera outbreaks.