In order to investigate the whole-cell proteomics and two-dimensional (2-D) gel maps of Aeromonas hydrophila isolated from the fish in this study, the proteomics analysis of A. hydrophila strain Zf-1 was performed using two-dimensional gel electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization timeof- flight mass spectrometry (MALDI-TOF/TOF-MS) techniques. A. hydrophila strain was prepared by shaking culture at 28 ℃ for 12 h, and the proteins of whole cell were extracted by using trichloroacetic acid (TCA)/acetone precipitation method. The results showed that 146 protein spots were observed in 2-DE gels performed with 18 cm pH3-10 IPG strips, and it shared matching profiles of about 81% protein expression spots while the 60 μg loading dose, and 7 cm pH3-10 IPG strips were used for isoelectric focusing (IEF) to 20 000 Vh. A total of 23 protein spots were further identified by the peptide mass fingerprinting (PMF) analysis, including Membrane lipoprotein, Molecular chaperones, 30S ribosomal protein S1, Elongation factor, Cytochrome C, etc; Furthermore, 11 enzymes of metabolism related proteins were found to be ATP F0F1 synthase subunit alpha, Dihydrolipoamide dehydrogenase, Enolase, Arginine deiminase, DNA-directed RNA polymerase subunit alpha, Glyceraldehyde-3- phosphate dehydrogenase, Carbamate kinase, Adenylate kinase, Uridine phosphorylase, Nitroreductase, and Peptidylprolyl isomerase. Analysis of protein functional classification with Gene Ontology (GO) of A. hydrophila based on molecular function showed that binding (17.1%), catalytic activity (14.3%), and transferase activity (11.4%) were responsible for the main biological functions. In conclusion, the proteomics analysis and 2-D map were constructed for A. hydrophila strain Zf-1 from the fish in this study, and the results will help to understanding the pathogenic mechanism and treatment of diseases caused by A. hydrophila from fish.