Antimicrobial agent residues are becoming an intractable environmental problem in soil, surface, and underground water. To obtain a broad profile of residues in animal wastewater and surface water, 24 animal wastewater, 8 animal farm effluent, 18 river water, and 8 pond water samples taken in Jiangsu in eastern China were monitored for enrofloxacin, ciprofloxacin, and florfenicol using solid phase extraction and high performance liquid chromatography/electrospray ionization–tandem mass spectrometry (HPLC/ESI–MS/MS) techniques. The results revealed that two antibacterials were detected simultaneously in 49.1% of samples, followed by three antibacterials (22.6%) and one antibacterial (22.6%). Up to 3.35, 5.93, and 2.10 μg L−1 for ciprofloxacin, 1.09, 4.24, and 0.50 μg L−1 for enrofloxacin, and 0.95, 2.40, and 2.84 μg L−1 for florfenicol were detected in animal farm-effluent, river, and pond water, respectively. The maximum concentrations of ciprofloxacin and enrofloxacin in animal wastewaters were 7.49 and 8.77 μg L−1, respectively. Furthermore, residue levels of ciprofloxacin and florfenicol showed at least two statistical differences between any two sampling areas or two animal farms. Enrofloxacin showed no statistical difference among the sampling areas and the animal farms.