Objectives: Veterinary practitioners’ working situation is both challenging and changing. They have higher levels of work-related stress and suicide risk than the general population. The proportion of women is increasing, and in Germany especially women and employed veterinarians are reported to be less satisfied than comparable subgroups of the general population. In this study we identified key factors associated with work and life satisfaction among veterinary practitioners in Germany. Design: Questionnaire-based cross- sectional survey. Setting: All veterinary practitioners registered in Germany in 2016. Participants: There were 2549 respondents, of whom 1930 met the inclusion criteria for further analysis. They had a median age of 37 and the majority of respondents were women (79.3 per cent). Almost two-thirds (63.8 per cent) worked as employed veterinarian. Primary outcomes: Importance of different job characteristics measured in 5-point Likert items, work satisfaction measured on a 5-point Likert item and life satisfaction measured in 11-point Likert items. Secondary outcomes: Facets such as satisfaction with leisure time, family life, health and standard of living, information on working conditions such as working time, income, as well as year of birth and other demographic data. Results: A ‘good working atmosphere’ was the most relevant job characteristic for all veterinary practitioners. Work satisfaction of employed practitioners is closely linked to satisfaction with their colleagues. This link is less pronounced for self-employed practitioners. A ‘reasonable salary’ was the second and ‘holidays and leisure time’ was the third most important job characteristics for employed practitioners. A ‘good working atmosphere’ and ‘family friendly arrangements’ were statistically significantly more important for women than for men, while a ‘reasonable salary’ was more important for men. Conclusions: Our results indicate strong associations between levels of work satisfaction and various work-related factors in subgroups of veterinary practitioners in Germany that reduce life satisfaction. The strength of some associations differs between men and women, as well as between self-employed and employed veterinarians. Outgoing students should be better prepared for the challenging working conditions that they face in veterinary practice. Salary levels should be improved and the working conditions adapted to the respective subgroups in order to increase work and life satisfaction.