Degree Name

Master of Science (MS)

Semester of Degree Completion

2017

Thesis Director

Kai Hung

Abstract

Proper treatment of municipal wastewater is important not only for prevention of eutrophication but also for public health concerns. One consideration for wastewater treatment is the presence of organisms carrying antibiotic resistance genes. If these organisms survive the wastewater treatment process and are released into the streams, they can pose significant risks to public health. In addition, their presence also raises concerns for lateral gene transfer events at the wastewater plant. The spread of antibiotic loci has a major impact in public health and clinical therapy because resistant organisms render conventional treatments of important illnesses difficult. Beta-lactam drugs are known to be the most prescribed antibiotics in United State for bacterial infections. However, their effectiveness is threatened by the spread of beta-lactamases, which produce products that destroy beta-lactam drugs. In this study, wastewater samples were taken at two stages — prior to primary treatment and right before release into the environment — for examination of the presence of 4 common variants of beta-lactamase using polymerase chain reactions. For comparison, freshwater samples from the stream both upstream and downstream of the wastewater efflux site were also collected and analyzed. Various DNA extraction methods were performed and compared for efficiency and quality. When combined with 16S rDNA control results, our data revealed that 3 of the 4 loci assayed were present in the influx wastewater sample but absent in both the efflux and stream water samples. This shows that for the sample we collected, the wastewater treatment process in the city of Charleston has reduced these antibiotic resistance loci to undetectable levels during the treatment process. Our data also showed that these resistant loci did not occur in untreated stream water.

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