|Mcmillan, Elizabeth A - University Of Georgia|
|Cho, Sohyun - University Of Georgia|
Submitted to: American Society for Microbiology General Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 6/2/2015
Publication Date: 6/2/2015
Citation: Hiott, L.M., Barrett, J.B., House, S.L., Woodley, T.A., Mcmillan, E., Cho, S., Jackson, C.R., Frye, J.G. 2015. Isolation and Analysis of Bacteria in Recreational Waters of the Chattahoochee River, Helen, GA. American Society for Microbiology General Meeting. May 30-June 2, 2015. New Orleans, Louisanna.
Technical Abstract: Helen is a tourism destination in the Appalachian Mountains. A popular activity during warm weather is tubing in the Chattahoochee River. This study was to determine the variety of bacteria in the Chattahoochee River in Helen, GA. Eight samples were collected during a 5km tubing trip down the Chattahoochee River on September 13, 2014. Every 10min, 100 mL of river water was collected in a sterile specimen cup and stored at 4°C. Duplicate 1 ml and 100 µL aliquots of each sample was applied to Petri film (3M, St. Paul, MN); 100 µL was plated to Tripticase Soy Agar with 5% sheep blood (BAP) (Fisher Scientific, Pittsburg, PA); 30 ml of each sample was also mixed with 30 ml Buffered Peptone Water (BPW) (Accumedia, Lansing, MI) for enrichment; all were incubated for 24h (all incubations are 37°C). 100 µl aliquots of each enrichment was spread onto BAP, CHROMagar ECC (Fisher), and CHROMagar O157 (DRG International, Springfield, NJ). Colonies from the BAP and both CHROMagar plates were streak isolated on BAP. For Salmonella, 1 ml from each enrichment was inoculated into selective enrichment broth [10 ml each Tetrathionate Broth (TET) (Fisher) and GN Broth, Hajna (Fisher)]. GN tubes were incubated 24h and TET 48h. 100 µl of GN and TET were transferred to 10 ml Rappaport-Vassiliadis R10 Broth (RV) and incubated for 24h.100 µl from the GN-RV broth and TET-RV broth was spread on Brilliant Green Sulfa (Fisher Scientific) and Xylose Lysine Tergitol-4 (Fisher) agar plates, incubated 24h, and suspect Salmonella colonies were streak isolated on BAP. Pure cultures on BAP were tested with KOH to determine Gram + or -. All bacteria were identified using the Vitek 2 Compact (BioMerieux, Durham, NC), using the appropriate Gram+ or Gram- panel. Coliform counts were 50 to 140/ml with one to two E. coli/ml detected in each sample. Overall, 265 bacteria were isolated, including 55 Aeromonas sp., 23 Acinetobacter sp., 18 Staphylococcus sp., 16 Psuedomonas sp., 11 E. coli and 5 Salmonella. Ten other genus were identified and 43 isolates could not be identified by Vitek.The majority of isolates recovered were naturally occurring bacteria from animals or the environment. Future studies will collect 1 L of water which will be concentrated by filtration before isolation of bacteria to increase our ability to recover Salmonella and other pathogens.