Screening of Biofilm-Forming Coral-Associated Bacteria using TC Plate By Crystal Violet Staining

Microbial communities are potentially very useful as indicators for water quality as they respond and be affected rapidly to environmental changes. Screening of biofilm-forming Coral-Associated bacteria can be used to study the natural and human factors affecting coasts and reefs. The objective of this experiment was to assess biofilm-forming in Coral-Associated bacterial isolates. Twelve strains of bacteria were grown in Tryptophan Soya Broth media and cultures were diluted into 100 folds, and then grown on a tissue culture (TC) plate. Crystal Violet staining was used to visualize biofilm growth. The procedure involved the addition of Crystal Violet solution, phosphate buffer saline (PBS) for washing and ethanol for fixation. The results show that 17% of the isolates displayed low biofilm growth patterns, while 25% of the isolates displayed medium biofilm growth patterns and 58% of the isolates displayed high biofilm growth patterns. When these results were compared with the appropriate controls, the Pseudomonas aerogenosa displayed high biofilm growth patterns while the negative control did not show any growth. In conclusion, Crystal Violet is a simple and easy method to evaluate the biofilm-forming differences among bacterial isolates, and in this experiment, it has been shown that formation of biofilm was very common in coral-associated bacteria. Keywords: Biofilm, Coral-Associated Bacteria, Tissue culture Plate, Microbial Communities https://doi.org/10.24897/acn.64.68.215