Microscopy and Fluorescence In-Situ Hybridization mini-study of four contaminant-degrading enrichment cultures and Biofilm formation study of the KB-1 and T3L cultures using confocal microscopy

Four environmentally-relevant and contaminant-degrading enrichment cultures (KB-1, ACT 3, T3L, and ORCH4) from the Edward’s Laboratory at the University of Toronto were studied via traditional microscopy, laser scanning confocal microscopy, and scanning electron microscopy. Also, general bacteria and archaea probes were tested for all these cultures to determine their suitability for continuous monitoring by Fluorescence In-situ Hybridization (FISH). For the culture ACT 3, a Dehalobacter specific probe (Dhb_442) was designed in-silico and experimentally tested in order to determine the morphology of the Dehalobacter species in this culture as well as in the sub-culture WL 1,2 DCA. Based on the preliminary results obtained, the relative abundance of Dehalobacter in ACT 3 and WL 1,2 DCA was 27 and 39%, respectively. Biofilm formation for the cultures KB-1 and T3L was also studied via laser scanning confocal microscopy. Biofilm was identified in both cultures; however, KB-1 produced larger and thicker biofilms in the time period studied.