Optical microscope, SEM (Scanning Electron Microscopy) and fluorescent microscope were used for qualitative and morphological studies of the attached biomass on PE (polyethylene) substratum under anaerobic condition. It was shown by the observation of optical microscope that the initial attachment of biomass began in crevices of the surface of PE. The shape and structure of the attached biofilm could be observed by SEM photographs, but species of bacteria were and methanogens were not classified. A large number of methanogenic bacteria were identified on the surface of PE substratum by fluorescence under 480nm of radiation. It was estimated that methanogenic bacteria was also related to initial attachment of biomass under anaerobic condition.