Pseudomonas alcaligenes is a nonfermenting Gram-negative bacillus and an unusual human pathogen belonging to the family Pseudomonadaceae. We describe the first case of a P. alcaligenes bloodstream infection in a premature neonate born at 33 weeks of gestation, who presented severe respiratory distress and was admitted to the Neonatal Intensive Care Unit at The Andes University Hospital in Mιrida, Venezuela. The strain was identified as P. alcaligenes by the matrix-assisted laser desorption ionization-time of flight mass spectrometry and 16S rRNA gene analysis. Antimicrobial susceptibility testing was carried out by E-test and broth microdilution methods. The isolate was resistant to ί-lactam/ί-lactamase inhibitor combinations, aztreonam, extended-spectrum cephalosporins, and gentamicin, but remained susceptible to carbapenems, fluoroquinolones, tobramycin, amikacin, minocycline, trimethoprim/sulfamethoxazole, and colistin. The neonate was successfully treated with meropenem. This case highlights the importance of P. alcaligenes as an emerging nosocomial neonatal pathogen.