Identifying bacteria from various environmental compartments is a common task in microbiology (e.g. infectiology, detection of water- and foodborne pathogens). Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) is a rapid and cost-effective method for bacterial species characterization. Most of the studies are focused on clinical isolates, but the appeal of this method is growing in environmental science. Here, we used MALDI-TOF MS as a screening tool to identify free living and plastic-associated bacteria of the lagoon of Toliara. Extracted macroplastic-associated bacteria have been cultured on different selective media and then identified as a set of 201 isolates by MALDI-Tof. The published dataset will provide insight into the pathobiome associated with floating plastic litter and help evaluate human and animal health risks caused by emerging and well-known pathogens species.