Laboratory Animals > Laboratory Animals Online First > 10.1258/la.2009.0080075

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Original Article

Charles River altered Schaedler flora (CRASF^®) remained stable for four years in a mouse colony housed in individually ventilated cages

Matthias Stehr ^1 *, Marina C Greweling ^2 3 * , Sabine Tischer ^1 4, Mahavir Singh ¹, Helmut Blöcker ¹, David A Monner ² and Werner Müller ^2 5

¹ Department of Genome Analysis, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany; ² Department of Experimental Immunology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany; ³ Present address: Division of Thoracic and Cardiovascular Surgery, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany; ⁴ Present address: Institute for Transfusion Medicine, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany; ⁵ Present address: Bill Ford Chair of Cellular Immunology, Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK; ^* The first two authors contributed equally to this work

Corresponding author: M C Greweling. Email: marina.greweling{at}helmholtz-hzi.de

As recommendations for specific pathogen-free housing change, mouse facilities need to re-derive their colonies repeatedly in order to eliminate specified bacteria or viruses. This paper describes the establishment of a new mouse facility using as starting point a small colony of CD-1 mice colonized with the Charles River altered Schaedler flora (CRASF^®) housed in individually ventilated cages (IVCs). The import of new strains was performed exclusively via embryo transfer using CD-1 mice as recipients. The integrity of the CRASF^® in caecum samples of the original CD-1 colony and of three inbred mouse lines imported into the colony was proven by a quantitative realtime polymerase chain reaction approach. Furthermore, we searched for bacterial contaminants in the gut flora using non-specific 16S rRNA primers. The bacterial sequences found were closely related to but not exclusively sequences of altered Schaedler flora (ASF) members, suggesting that the ASF is heterogeneous rather than restricted to the eight defined bacteria. Moreover, no pathogens were found, neither using the non-specific 16S rRNA primers nor in routine quarterly health monitoring. As one effect of this defined gut flora, interleukin-10 knockout mice are devoid of colitis in our facility. In conclusion, our approach building up a mouse facility using foster mothers and embryo transfer as well as a strict barrier system and IVCs is suitable to maintain a colony free from contaminating bacteria over the long term. CRASF^® remained stable for seven mouse generations and was efficiently transferred to the imported mouse strains.

Key Words: Altered Schaedler flora • IVC • realtime PCR • 16S rRNA

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