What Is Contaminating Streams in the Buffalo Creek Watershed?

The state of Pennsylvania is surrounded by numerous streams and bodies of water, some as easily accessible as being in your backyard. However, the majority of these waterways are inadequate for recreational activities due to nutrient pollution, bacteria and pathogen contamination, and sediment pollution. Despite waterways being assessed for impairment, the general public remains to have access to these bodies of water with no warning of the water’s quality. After collecting and evaluating water samples from 13 locations in the Buffalo Creek Watershed, five times over a period of 30 days for the summer of 2022 and 2023, we have determined that the majority of the streams demonstrate high bacteria concentrations from fecal pollution. It is hypothesized that this fecal pollution is derived from farming practices as Union County is rural.

From the water samples collected, filtration for DNA extraction was done using Millipore Express PLUS 0.22 μm filters. The addition of 100 μL of 1.2 mg/L Salmon Sperm DNA to filters was done before using Qiagen's DNeasy PowerSoil Kit for DNA extraction. The collection of animal fecal matter was also included in this research and involved samples of pig, cow, goose, chicken, dog, deer, human, and horse fecal samples collected within the area. From these samples, DNA extraction was performed using Zymo Research’s Quick-DNA Fecal/Soil Microbe Miniprep Kit. After DNA extraction was done for water and fecal samples, a Polymerase Chain Reaction (PCR) was performed for each DNA extraction to make millions of copies of the DNA of interest. Gel electrophoresis was then performed with the PCR samples of animal extracted DNA to determine the specificity and sensitivity of our animal primers; CowBACB2, CowM3, Pig-2, Avian GFD, Goose CG1, Deer EF447, Dog DG3, Human HF183, and Horse HoF597F.

Once we have established the specificity and sensitivity of our animal primers to the correct animal DNA, we can move towards determining what animal fecal matter is polluting the streams. Using the data of the animal DNA present in specific water samples along with the extracted data from our county’s Nutrient Management Plans (NMPs), we can use ArcGIS to map and pinpoint potential sources of contamination.