Determining Sources of Fecal Pollution in a Rural Virginia Watershed with Antibiotic Resistance Patterns in Fecal Streptococci

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Applied and Environmental Microbiology, December 1999, p. 5522-5531, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Determining Sources of Fecal Pollution in a Rural Virginia Watershed with Antibiotic Resistance Patterns in Fecal Streptococci

Charles Hagedorn,^* Sandra L. Robinson, Jennifer R. Filtz, Sarah M. Grubbs, Theresa A. Angier, and Raymond B. Reneau Jr.

Department of Crop and Soil Environmental Sciences, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061

Received 4 June 1999/Accepted 12 September 1999

Nonpoint sources of pollution that contribute fecal bacteria to surface waters have proven difficult to identify. Knowledgeof pollution sources could aid in restoration of the water quality,reduce the amounts of nutrients leaving watersheds, and reducethe danger of infectious disease resulting from exposure to contaminatedwaters. Patterns of antibiotic resistance in fecal streptococciwere analyzed by discriminant and cluster analysis and used toidentify sources of fecal pollution in a rural Virginia watershed.A database consisting of patterns from 7,058 fecal streptococcusisolates was first established from known human, livestock, andwildlife sources in Montgomery County, Va. Correct fecal streptococcussource identification averaged 87% for the entire database andranged from 84% for deer isolates to 93% for human isolates. Tofield test the method and the database, a watershed improvementproject (Page Brook) in Clarke County, Va., was initiated in 1996.Comparison of 892 known-source isolates from that watershed againstthe database resulted in an average correct classification rateof 88%. Combining all animal isolates increased correct classificationrates to $>=$ 95% for separations between animal and human sources.Stream samples from three collection sites were highly contaminated,and fecal streptococci from these sites were classified as beingpredominantly from cattle (>78% of isolates), with small proportionsfrom waterfowl, deer, and unidentified sources ( $approx$ 7% each). Basedon these results, cattle access to the stream was restricted byinstallation of fencing and in-pasture watering stations. Fecalcoliforms were reduced at the three sites by an average of 94%,from prefencing average populations of 15,900 per 100 ml to postfencingaverage populations of 960 per 100 ml. After fencing, <45% offecal streptococcus isolates were classified as being from cattle.These results demonstrate that antibiotic resistance profilesin fecal streptococci can be used to reliably determine sourcesof fecal pollution, and water quality improvements can occur whenefforts to address the identified sources aremade.

^* Corresponding author. Mailing address: Department of Crop and Soil Environmental Sciences, VPI & SU, Smyth Hall, Blacksburg,VA 24061-0404. Phone: (540) 231-4895. Fax: (540) 231-3431. E-mail:chagedor{at}vt.edu.

Applied and Environmental Microbiology, December 1999, p. 5522-5531, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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