Neonatal sepsis refers to the bacterial bloodstream infections of the newborn during the neonatal period as usually the first twenty-eight days of life. The current study was done in the laboratories of AL-Batool Teaching Hospital for Gynecology and Pediatrics in Baqubah, Diyala Governorate, including 140 blood specimens collected from the neonates admitted to the hospital with suspected sepsis, the ages of the both groups was ranged from 1 day to 28 days. Out of the total cultured samples, 32.14% (45 of 140) were positive and 67.86% (95 of 140) were negative blood culture. 45 of 140 samples were negative to the blood culture chosen as control group. The results showed highest isolates were Coagulase Negative Staphylococcus (CoNS) 19 (42.2%), followed by Staphylococcus aureus 11 (24.5%), Escherichia coli 4 (8.8%), Klebsiella pneumonia 4 (8.8%), Acinetobacter baumannii 3 (6.7%), Group B Streptococcus (GBS), and Pseudomonas aeruginosa 2 (4.5%). Ceftazidime antibiotic has the highest resistance percentage followed by CIP, CXM, AMP, NA, C, CD, and CL among the studied bacterial isolates. Biofilm formation of isolates showed all bacterial isolates of K. pneumonia, A.baumannii, P.aeruginosa, and GBS by 100% can form biofilm, while the isolates of S.aureus, CoNS and E.coli were 6 (53.55%), 7 (36.84%), and 2 (50%) biofilm forming, respectively. These biofilm-forming isolates exhibited high resistance to AK, AMC, TM, CTX, NE, VAN, COT, CL, CAZ, AMP, NA, C, and CIP.
