Characterization of Protozoa Transport and Occurrence of Chlorinated-Ethene Reducer Bacteria in Subsurface Environments

Abstract

This dissertation contains the results of two different projects. The first one is a study of the transport of protozoa pathogens Cryptosporidium parvum and Encephalitozoon intestinalis in soils. The aim of this project was to investigate the movement and retention mechanisms of these microorganisms in natural porous media. The work determined that in the case of C. parvum, the retention was primarily produced by straining and in the case of E. intestinalis the main retention mechanism was attachment. The results of C. parvum lysimeter experiment compared to the results from the 7 cm column experiments suggest that retention is proportional to the length of the column. The second study evaluated the Polymerase Chain Reaction (PCR) as a tool to identify dechlorinating bacteria in groundwater contaminated with chloroethenes. The target DNA regions to identify these microorganism were the 16s rDNA specific for dehalococcoides sp. and Desulfuromonas and DNA sequences coding for the reductive dehalogenase enzymes pceA, tceA, bvcA and vcrA. Bacteria able to transform PCE into DCE were detected in all groundwater samples. Bacteria able to transform VC into ethene were found only in one of the samples. This study shows that PCR analysis of 16s rDNA and reductive dehalogenase gene sequences together with microcosm results are useful tools to analyze the populations of reductive dechlorinators and their activity in a given site.