AEM Accepts, published online ahead of print on 22 June 2007
This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ufnar, J. A.
Right arrow Articles by Ellender, R. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ufnar, J. A.
Right arrow Articles by Ellender, R. D.
Agricola
Right arrow Articles by Ufnar, J. A.
Right arrow Articles by Ellender, R. D.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol. doi:10.1128/AEM.00319-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Development of a Swine-Specific Fecal Pollution Marker Based on Host Differences in Methanogen mcrA Genes

Jennifer A. Ufnar*, David F. Ufnar, Shiao Y. Wang, and R. D. Ellender

Department of Biological Sciences, The University of Southern Mississippi, Hattiesburg, MS 39406; Department of Geography and Geology, The University of Southern Mississippi, Hattiesburg, MS 39406

* To whom correspondence should be addressed. Email: jennifer.ufnar{at}usm.edu.


arrow
Abstract

The goal of this study was to evaluate methanogen diversity in animal hosts to develop a swine-specific archaeal molecular marker for fecal source tracking in surface waters. Phylogenetic analysis of swine mcrA sequences compared to mcrA sequences from the feces of five animals (cow, deer, sheep, horse, and chicken) and sewage showed four distinct swine clusters, with three swine-specific clades. From this analysis, six sequences were chosen for molecular marker development and initial testing. Only one mcrA sequence (P23-2) showed specificity for swine and was used for environmental testing.

PCR primers for the P23-2 clone mcrA sequence were developed and evaluated for swine-specificity. The P23-2 primers amplified products in P23-2 plasmid DNA (100%), pig feces (84%), and swine waste lagoon surface water samples (100%), but did not amplify a product in 47 bacterial and archaeal stock cultures, 477 environmental bacterial isolates, sewage, and water samples from a bovine waste lagoon and polluted creek. Amplification was observed in only 1 sheep out of 260 human and non-swine animal fecal samples. Sequencing of PCR products from pig feces demonstrated 100% similarity to pig mcrA sequence from clone P23-2. The minimal amount of DNA required for the detection was 1 pg for P23-2 plasmid, 1 ng for pig feces, 50 ng for swine waste lagoon surface water, 1 ng for sow waste influent, and 10 ng for lagoon sludge samples. Lower detection limits of 10-6 g of wet pig feces in 500 mL of PBS and 10-4 g of lagoon waste in estuarine water were established for the P23-2 marker. This study was the first to utilize methanogens for the development of a swine-specific fecal contamination marker.




This article has been cited by other articles:

  • Lu, J., Santo Domingo, J. W., Hill, S., Edge, T. A. (2009). Microbial Diversity and Host-Specific Sequences of Canada Goose Feces. Appl. Environ. Microbiol. 75: 5919-5926 [Abstract] [Full Text]  
  • Lamendella, R., Santo Domingo, J. W., Yannarell, A. C., Ghosh, S., Di Giovanni, G., Mackie, R. I., Oerther, D. B. (2009). Evaluation of Swine-Specific PCR Assays Used for Fecal Source Tracking and Analysis of Molecular Diversity of Swine-Specific "Bacteroidales" Populations. Appl. Environ. Microbiol. 75: 5787-5796 [Abstract] [Full Text]  
  • Marti, R., Dabert, P., Pourcher, A.-M. (2009). Pig Manure Contamination Marker Selection Based on the Influence of Biological Treatment on the Dominant Fecal Microbial Groups. Appl. Environ. Microbiol. 75: 4967-4974 [Abstract] [Full Text]  
  • Mieszkin, S., Furet, J.-P., Corthier, G., Gourmelon, M. (2009). Estimation of Pig Fecal Contamination in a River Catchment by Real-Time PCR Using Two Pig-Specific Bacteroidales 16S rRNA Genetic Markers. Appl. Environ. Microbiol. 75: 3045-3054 [Abstract] [Full Text]  
  • Ufnar, J. A., Wang, S. Y., Ufnar, D. F., Ellender, R. D. (2007). Methanobrevibacter ruminantium as an Indicator of Domesticated-Ruminant Fecal Pollution in Surface Waters. Appl. Environ. Microbiol. 73: 7118-7121 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»