NR AWJN
AU Maluquer de Motes,C.; Simon,S.; Grassi,J.; Torres,J.M.; Pumarola,M.; Girones,R.
TI Development of methods for the analysis of the presence and stability of prions in wastewater
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions EPI-14
PT Konferenz-Poster
AB Experimental approaches on the presence and behaviour of prion proteins in the wastewater generated in abattoirs, are required in order to provide experimental data for the evaluation of sanitary risks related with the dissemination of the contamination and development of good management practices. Some quantitative risk assessment studies have hypothesized a possible transmission of prion diseases through sewage and sludge from abattoirs where TSE cases have been identified, especially BSE. As only post-mortem tests are available and all slaughtering processes are finished when the TSE case is confirmed, some SRM infectivity could enter the sewer inducing a prion contamination. Furthermore, prions are highly resistant, markedly insoluble and amphipathic. The objective of this work was to develop a methodology to deal with prions in slaughterhouse sewage, providing experimental tools to evaluate their possible presence and their persistence in these matrices. Thus, different slaughterhouse sewage samples were spiked with either scrapie or BSE brain homogenates and brain tissues in suspension were recovered by centrifugation. The corresponding pellet was resuspended, PK-treated and concentrated using the Bio-Rad TSE Purification kit with some modifications, and finally analyzed by Western blot. The designed procedure has been able to detect 2ug of BSE- and 0.5ug Scrapie-affected tissue for 30ml of sewage sample analyzed. This procedure was applied to the analysis of 17 sewage samples collected in slaughterhouses processing cows of ages older than 24 months. Eight of these samples were collected in slaughterhouses one day after the identification of a TSE positive case between the processed animals. None of the samples was found to be positive forC PrPsc. The stability of PrPsc in wastewater was experimentally studied at 20 deg C. Persistence curves for either scrapie or BSE were obtained in the experiments analyzing raw wastewater samples and in samples collected after a primary treatment. In conclusion, the developed methodology for detection of PrPsc in slaughterhouse sewage may represent a sensitive technique useful for detecting either Scrapie or BSE associated PrPsc wastewater and as a tool for the study and the control of the dissemination of prions in the environment.
AD C. Maluquer de Motes, R. Girones: Department of Microbiology, University of Barcelona, Diagonal 645, 08028 Barcelona, Spain; S. Simon, J. Grassi: CEA, Service de Pharmacologie et d'Immunologie, CEA/Saclay, 91191 Gif sur Ivette, France; J.M. Torres: Centro de Investigación en Sanidad Animal (CISA-INIA), Ctra. De Valdeolmos a El Casar, Valdeolmos, Madrid, Spain; M. Pumarola: PRIOCAT laboratory, CReSA, Veterinary Faculty, Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallés), 08193 Barcelona, Spain
SP englisch
PO Italien