Methods of isolation and identification of pathogenic and potential pathogenic bacteria from skins and tannery effluents

Currently there is no standard protocol available within the leather industry to isolate and identify pathogenic bacteria from hides, skins or tannery effluent. This study was therefore carried out to identify simple but effective methods for isolation and identification of bacterial pathogens from the effluent and skins during leather processing. Identification methods based on both phenotypic and genotypic characteristics were investigated. Bacillus cereus and Pseudomonas aeruginosa were used as indicator bacteria to evaluate the isolation and identification methods. Decontaminated calfskins were inoculated with a pure culture of the above mentioned bacterial species followed by a pre-tanning and chromium tanning processes. Effluent samples were collected and skins were swabbed at the end of each processing stage. Bacterial identification was carried out based on the phenotypic characteristics; such as colony appearance on selective solid media, cell morphology following a standard Gram-staining and spore staining techniques, and biochemical reactions, e.g., the ability of a bacterial species to ferment particular sugars and ability to produce certain enzymes. Additionally, an identification system based on bacterial phenotypic characteristics, known as Biolog® system was applied. A pulsed-filed gel electrophoresis (PFGE) method for bacterial DNA fingerprinting was also evaluated and used for the identification of the inoculated bacteria. The methods described in the study were found to be effective for the identification of pathogenic bacteria from skins and effluent.