Back ground: Pre-analytical activities persist to pose significant amount of uncertainties in clinical laboratories causing adverse impacts on patient health and the entire healthcare system. Venous blood sample (VBS) processing that has many error prone activities is one of the pre-analytical procedures performed in the laboratory. The objective of our survey was to identify, in Ethiopian Hospital laboratory setup , the major activities in the processing of VBS in which undesirable practices that may result in errors are executed. Methods: We have conducted institution based cross-sectional survey supplemented with non-participatory type observational study from February 2012 to September 2012 in laboratories of three governmental hospitals of Gamo Gofa zone, Southern Ethiopia. Pre-tested questionnaire and check list were used for data collection. Analysis of the data was performed using Medcalc® version 12.1.4 software. Results: A total of 19 laboratory professionals working in the three governmental hospitals were included in this survey. The activities possessing highest proportions of undesirable practices were related to establishment and adherence to serum/plasma/whole blood rejection criteria, measures taken when produced serum/plasma is too small for analysis, speed and duration of centrifugation. Low proportion of undesirable practices were found in activities related to capping test tubes before centrifugation, maximum allowed time before analysis of unpreserved serum/plasma/whole blood samples and balancing during centrifugation. None of the socio-demographic and background information of participants we assessed was associated with undesirability of VBS processing activities. Conclusion: From this study, we concluded that the VBS processing in the laboratories involved many undesirable practices that might lead to erroneous results. We identified that the gearing problem to the undesirable practices was absence of laboratory documents regarding VBS processing activities. Therefore, establishment and strict adherence to laboratory documents for every activity in VBS processing by every laboratory personnel would avoid many of the undesirable practices.