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Applied and Environmental Microbiology, October 2003, p. 6311-6315, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.6311-6315.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Teschoviruses as Indicators of Porcine Fecal Contamination of Surface Water

Miguel Angel Jiménez-Clavero,1 Carlos Fernández,2 José Antonio Ortiz,2 Javier Pro,2 Gregoria Carbonell,2 José Vicente Tarazona,2 Neftalí Roblas,1 and Victoria Ley1*

Departamento de Biotecnología,1 Departamento de Medio Ambiente, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), 28040 Madrid, Spain2

Received 27 February 2003/ Accepted 2 July 2003

Teschoviruses specifically infect pigs and are shed in pig feces. Hence, their presence in water should indicate contamination with pig fecal residues. To assess this hypothesis, we have developed a real-time reverse transcriptase PCR (RT-PCR) method that allows the quantitative detection of pig teschovirus (PTV) RNA. The method is able to detect 92 fg of PTV RNA per ml of sample. Using this method, we have detected the presence of PTV RNA in water and fecal samples from all pig farms examined (n = 5). Feces from other animal species (cattle, sheep, and goats) were negative in this test. To compare the PTV RNA detection method with conventional chemical determinations currently in use for evaluation of water contamination, we analyzed water samples collected downstream from a pig slurry spillage site. We have found a positive correlation within both types of determinations. The sensitivity of the PTV detection assay was similar to that achieved by unspecific organic matter determination and superior to all other conventional chemical analyses performed. Furthermore, the new method is highly specific, revealing the porcine origin of the contamination, a feature that is lacking in currently available methods for the assessment of water contamination.

* Corresponding author. Mailing address: Departamento de Biotecnología, Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria, Ctra. Coruña Km 7.5, 28040 Madrid, Spain. Phone: 3491 3471497. Fax: 3491 3572293. E-mail: ley{at}inia.es.

Applied and Environmental Microbiology, October 2003, p. 6311-6315, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.6311-6315.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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