Background Determining burden of pulmonary tuberculosis (PTb) may be associated with success of directly observed theraphy (DOT) services in high PTb areas, such as Gambella, Ethiopia. Objective To assess burden of sputum smear positive SSP-PTb reports in Gambella Regional Hospital, in two consecutive years: mid 2013 to 2015. Methods From March 2013 to February 2015, PTb suspected patients' sputum Acid Fast Bacilli test results were collected from laboratory registration book of Gambella Regional Hospital, western Ethiopia. Results Of the total 3064 results, the proportions of males and individuals whose age is between 16 and 25 years were 1823 (59.5%) and 1046 (34.1%), respectively. Majority of samples were coming from newly PTb suspected patients (2587, or 84.4%); and of them, SSP-PTb cases were 9.9%. The overall SSP-PTb cases decreased by 2.1% in March 2014 to February 2015 as compared to March 2013 to Feburary 2014. Meanwhile, AFB smear negative cases were significantly associated with being male [1.384 (1.064–1.801)] and being of age group 16–25 years [1.517 (1.007–2.287)]. Conclusion In Gambella region, although the distribution of AFB smear results had no significant yearly variations and the overall burden of SSP cases was decreased, still a lot of work needs to be undertaken on the PTb prevention and control program.