Abstract: Ovarian hydrobursitis syndrome represents a major reproductive problem in dromedary. The aim of the present study was to investigate the causative agents of such syndrome. A total of sixty one non-repetitive isolates (twenty from infection site and 41 from water sources used by camels for drinking) were collected from Al Ahsa province, eastern region of Saudi Arabia. Vitek 2 compact automated system was used to identify the isolates, to determine minimum inhibitory concentration of twenty antibiotics and to explore the antibiogram of certain isolates. Polymerase chain reaction was used to amplify genes encoding extended spectrum β-lactamases (ESBL). Escherichia coli (E. coli) isolates were the most common where 17 isolates were collected from affected animals while three isolates were collected from the water sources. In addition, two Klebsiella pneumoniae and one Pseudomonas aeruginosa isolates were collected from infection sites. Moreover, three ESBL producing E. coli were identified (two isolates from affected animals and one isolate from the water sources). These three isolates harbored blaTEM while blaSHV and blaCTX-M were not detected. The present study showed that additional infective agents could be associated with ovarian hydrobursitis. In conclusion, strict infection control measures should be applied to prevent spreading of such multidrug resistant pathogens.