Objective: Marine bacteria are a rich source of potentially useful antimicrobial molecules. The purpose of the study is to explore the diversity of bacteria with antimicrobial activity isolated from Siganus fuscescens gastrointestinal tract collected from Naozhou Island (20 degrees 52' N-20 degrees 56' N 110 degrees 33' E-110 degrees 38' E), Leizhou Bay, South China Sea., Methods: We isolated bacteria from the gastrointestinal tract of fish sample using classical culturing technique, and determined antimicrobial activities of the isolates by Oxford cup method. We investigated diversity of antimicrobial isolates using phylogenetic comparative analysis of 16S rRNA gene sequences., Results: According to the results of morphological observation and part of physiological and biochemical experiments, we isolated 68 strains from fish gastrointestinal tract. Among them, 19 strains with antimicrobial activities were acquired (27.9% of the isolates) and represented 19 different species, belonging to 12 genera (Rothia, Micrococcu, Brachybacterium, Brevibacterium, Psychrobacter, Paracoccus, Cobetia, Citrobacter, Pseudomonas, Bacillus, Lactobacillus and Staphylococcus) of 11 families (Microbacteriaceae, Dermabacteraceae, Brevibacteriaceae, Moraxellaceae, Rhodobacteraceae, Halomonadaceae, Enterobacteriaceae, Pseudomonadaceae, Bacillaceae, Lactobacillaceae and Staphylococcaceae) in three phyla (Actinobacteria, Proteobacteria and Firmicutes). Eight strains (42.1%), 7 strains (36.8%) and 4 strains (21.1%) were belonging to the phylum Firmicutes, Proteobacteria and Actinobacteria, respectively. The phylogenetic distance matrix results suggested that there were obvious genetic divergences between the majority of strains with antimicrobial activity and their phylogenetically most closely related typical strain, due to 16S rRNA gene sequences similarities ranging from 96.2 to 99.9%. In addition, 4 strains (ZJHD2-31, ZJHD5-23, ZJHD2-58 and M26) could represent potential new species, and identification of the novel strain M26 has been published in Antonie van Leeuwenhoek., Conclusion: There are abundant diversity for bacteria with antimicrobial activity and potentially more new species of microorganism in Siganus fuscescens gastrointestinal tract collected from Naozhou Island.