To detect the tetrodotoxin (TTX) of Takifugu niphobles and its symbiotic bacteria, three individuals of T. niphobles were collected from the sea near County Dongshan, Zhangzhou, Fujian Province, China. The collected fishes T. niphobles owned some typical morphological features, such as a lot of white dots spreading all over the dorsal body, each of oval black patches is at the base of dorsal fin or behind pectoral fin in each body side, each fins showing light yellow color. According to the test results of liquid chromatography coupled with mass spectrometry (LC-MS/MS), the amount of TTX in ovary, liver and muscle tissue contain 56.247, 0.380 and 0.118 μg/g respectively. Based on the 16S rDNA sequence identity and phylogenetic analysis with known bacterial species in GenBank database, one bacterial strain Listonella sp. from intestines, Enterobacter sp. from muscle and Vibrio crassostreae from ovary, two bacteria Acinetobacter sp. from liver, were isolated in this study. According to the LC-MS/MS result, three bacteria can produce TTX except Vibrio crassostreae and one bacterial strain Acinetobacter sp.. 10¹³ CFU of Acinetobacter lwoffii, Enterobacter sp. and Listonella sp. can produce 0.855, 0.318, and 0.298 ng TTX, respectively. This is the first report that the bacteria of Listonella and Acinetobacter isolated from puffer fish can produce TTX, our research further enriched the catalogue of TTX-producing bacteria.