Cellular respiration is a fundamental feature of metabolic activity and oxygen consumption can be considered as a reliable indicator of bacterial aerobic respiration, including for facultative anaerobic bacteria like E. coli. Addressing the emerging global health challenge of antimicrobial resistance, we performed antimicrobial susceptibility testing using the bacterial oxygen consumption rate (OCR) as a phenotypic indicator. We demonstrated that microbial exposure to antibiotics showed systematic OCR variations, which enabled determining minimum inhibitory concentrations for three clinically relevant antibiotics, ampicillin, ciprofloxacin, and gentamicin, within a few hours. Our study was performed by using photoluminescence-based oxygen sensing in a microchamber format, which enabled reducing the sample volume to a few hundred microliters. OCR modeling based on exponential bacterial growth allowed estimating the bacterial doubling time for various culture conditions (different types of media, different culture temperature and antibiotic concentrations). Furthermore, correlating metabolic heat production data, as obtained by nanocalorimetry in the same type of microchamber, and OCR measurements provided further insight on the actual metabolic state and activity of a microbial sample. This approach represents a new path towards more comprehensive microbiological studies performed on integrated miniaturized systems.