Abstract: Objective: The intent of this study was to characterize the potential bacterial contamination before and after sterilization in a limited resource setting and re-assess good surgical practices. This included the monitoring of antimicrobial susceptibility of recovered isolates, which have bearing on optimal antimicrobial use. Materials and Methods: Samples were collected from three surgical sites on the patient (two before cutting and one after), surgeon’s hand and pooled sample from surgical instruments in sterilization process. Antimicrobial susceptibility was performed on recovered bacteria. Descriptive analyses were done to compare between isolate recovery between sites and susceptibility findings. Results: Dogs enrolled in the study ranged in age from 0.5-6 years (mean 2.4). Bacterial isolates were recovered from 28 (93%) dogs; 17 (57%) surgeon’s hands and 8 (27%) pooled surgical equipment swabs. Eighty two unique bacterial isolates were recovered from 150 collected samples. Of the 82 isolates, 18 were identified as Staphylococcus sp. and 64 were identified as Bacillus sp. and Staphylococcus sp., were generally susceptible to amoxicillin/clavulanic acid, cephalexin, chloramphenicol and gentamycin. Thirteen different antimicrobial resistance patterns were observed for Staphylococcus sp. Four isolates were pan-sensitive. About 10 of 18 isolates were resistant to three of more classes of antibiotics. Conclusion: Reproductive surgical sterilization is an important tool in efforts to control pet overpopulation, even in limited resource settings. With ongoing concerns about emerging antibiotic resistance, implementation of optimum aseptic technique is needed. Findings from this study support standard and appropriate surgical protocols in dog control sterilization in limited resource settings which supports optimal antibiotic use policies.