An ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS / MS)method was developed for the simultaneous determination of 34 fluoroquinolones，sulfonamides and tetracyclines in fresh sheep manure，fresh cow manure，fresh pig manure，fresh chicken manure，treated pig manure and treated chicken manure. Livestock manure were extracted with methanol and Na2EDTA-Mcllvaine extraction solution for 2 times，treated with ultrasonic cleaner for 10 min，centrifuged for 5 min，purified by solid phase extraction，and determined by LC-MS / MS external standard method. The results showed that there was a good linear relationship between the 34 antibiotics in fresh cow manure，fresh sheep manure，fresh pig manure，treated pig manure，fresh chicken manure and treated chicken manure. The recovery rates of standard addition were between 64.6% and 117.9%. This method can meet the analysis requirements of 34 antibiotics content of fluoroquinolones，sulfonamides and tetracyclines in livestock manure.