Abstract: In this study, investigation of the presence of Staphylococcus and determination of its prevalence and distribution, identification of Staphylococcus species and determination of their prevalence and distribution and characterization of the isolates in order to determine their ability in synthesizing coagulase from raw bovine milk samples were conducted from October 2008-April 2009 in Debre Zeit, Ethiopia. About 200 raw bovine milk samples consisting of 100 buckets milk of farms and 100 tanks milk of milk collection centers were analyzed. The identification results showed 33 and 46% prevalence of Staphylococcus in buckets milk and tanks milk, respectively with an overall prevalence of 39.5% (79/200). Comparison of the prevalence of Staphylococcus in raw bulk milk samples showed a relatively higher prevalence in tanks milk (46%) than buckets milk (33%). However, this difference was not statistically significant (p>0.05). The 33 isolates identified as staphylococci from samples of buckets milk were tested for species assignment. They were grouped into S. aureus with 8 (8%) isolates, S. intermedius with 6 (6%) isolates, S. hycius with 6 (6%) isolates and Coagulase Negative Staphylococci (CNS) with 13 (13%) isolates. The 46 staphylococci isolates were grouped into S. aureus with 10 (10%) isolates, S. intermedius with 11 (11%) isolates, S. hycius with 6 (6%) isolates and CNS with 19 (19%) isolates. There was no significant difference (p>0.05) among these proportion of isolates in both buckets and tanks milk. All the isolates were tested for the production of coagulase to determine their pathogenicity. Comparison of the prevalence of Coagulase Positive Staphylococci (CPS) showed a relatively higher CPS prevalence in tanks milk (27%) than buckets milk (20%). However, this difference was not statistically significant (p>0.05). The high level of Staphylococcus isolate found raw milk samples in the present study represent a poor quality and public health risk to the consumer. Hence, raw milk intended for human consumption must be subjected to pasteurization or heat treatment at least equivalent to pasteurization temperature order to guarantee the quality of these highly popular products in Debre Zeit in order to decrease the risk of staphylococcal food poisoning.