Stabilization of microorganisms for in situ degradation of toxic chemicals

Methods for large-scale microencapsulation of bacteria and nutrients into microbeads with small enough diameters to travel through aquifers have been developed at the University of Idaho. Both free and immobilized cells of Flavobacterium ATCC 39723, a gram-negative aerobe that degrades various chlorinated phenols, into aquifer microcosms, through which pentachlorophenol (PCP)-contaminated groundwater flowed at in situ flow rates. Aquifer samples were collected with an auger from three wells at the University of Idaho Ground Water Research Site, and packed into 24 columns. Some sterile columns were also prepared, by irradiation at the Washington State University Radiation Center. In some of the columns the free Flavobacterium cells were mixed with the aquifer material before packing the columns. In others, agarose-microimmobilized Flavobacterium were mixed into the aquifer material. The effluent from each column was collected daily for 170 days and analyzed by UV spectroscopy or HPLC for remaining PCP. There were no statistically significant differences between the degradation rates of free or encapsulated Flavobacterium in sterile or native aquifer material as tested in these experiments. This work has shown at the lab scale that encapsulated PCP-degrading Flavobacterium were able to survive under conditions of starvation, predation, and lack of water