The invention discloses automatic monitoring equipment for multi-strain water biotoxicity. The automatic monitoring equipment comprises a PCL unit, a man-machine interaction unit, a flow path control unit, an anabiosis preservation unit, a sampling quantifying unit and a reaction detection unit, wherein the PCL unit, the man-machine interaction unit, the flow path control unit, the anabiosis preservation unit, the sampling quantifying unit and the reaction detection unit are all arranged inside a frame; the PCL unit is connected with the man-machine interaction unit, the flow path control unit and the reaction detection unit; the flow path control unit is connected with the anabiosis preservation unit, the sampling quantifying unit and the reaction detection unit. The invention further discloses a monitoring method for the multi-strain water biotoxicity. The automatic monitoring equipment has the advantages of multi-strain applicability, whole-journey detection, high measurement precision, short measurement time, small bacterial liquid amount, no cross interference, small maintenance amount and the like, and requirements of on-line environment monitoring can be met.