Identification of Microorganisms for the Bioremediation of Nitrate and Manganese in Minnesota Water

Abstract

Bioremediation is a way to safely and cost-effectively remove contaminants using living organisms. In this thesis, microorganisms capable of remediating two pollutants, nitrate and manganese, were identified using culture-dependent and –independent approaches. Nitrate in agricultural wastewater can lead to algal blooms and eutrophication. Edge-of-field woodchip bioreactors are a promising approach to prevent nitrate in wastewater from reaching surface waters by utilizing microbial denitrification to remove nitrate from the system. However, woodchip bioreactors experience low efficiency under cold temperatures, so one strategy to enhance bioreactors in the early spring involves bioaugmentation, or inoculating the bioreactors with cold-adapted denitrifying microorganisms. In order to identify a cold-adapted denitrifier for bioaugmentation, microorganisms were isolated from field woodchip bioreactors and subjected to denitrification testing under cold temperatures, measuring nitrate, nitrite, ammonium, nitrous oxide and dinitrogen gas, as well as whole genome sequencing to identify the presence of genes involved in denitrification and other important microbial processes. Based off of these results, two strains, Microvirgula sp. BE2.4 and Cellulomonas sp. WB94 were recommended for bioaugmentation. In part two, manganese was addressed. High levels of manganese in drinking water can cause health problems, and common treatment methods require cost-intensive chemicals, conditions and maintenance. In this study, a novel algae bioreactor was established to remove manganese from water. In this bioreactor, the algae provided fixed carbon for manganese-oxidizing microorganisms that oxidized the dissolved manganese, precipitating it out of solution. Using a culture-dependent approach, manganese-oxidizing bacteria and fungi were isolated from an environmental sample, including known oxidizers Bosea, Pseudomonas, Plectosphaerella and Phoma and some not previously known to oxidize manganese such as Aeromonas, Skermanella, Ensifer and Aspergillus. A culture-independent approach was also employed to determine how abundant the isolated manganese-oxidizing bacteria are in an actively oxidizing environmental sample. Using nitrate and manganese as examples, this thesis identified useful microorganisms involved in remediation and demonstrated how microorganisms can be utilized to effectively remove pollutants from the environment.