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Applied and Environmental Microbiology, April 2005, p. 2086-2094, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.2086-2094.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Explorative Multifactor Approach for Investigating Global Survival Mechanisms of Campylobacter jejuni under Environmental Conditions

Birgitte Moen,^1,2* Astrid Oust,^1,2 Øyvind Langsrud,² Nick Dorrell,³ Gemma L. Marsden,^4, Jason Hinds,⁴ Achim Kohler,² Brendan W. Wren,³ and Knut Rudi^2*

Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences,¹ Matforsk, Norwegian Food Research Institute,Ås, Norway,² Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine,³ Department of Cellular and Molecular Medicine, St. George's Hospital Medical School, London, United Kingdom⁴

Received 15 July 2004/ Accepted 19 October 2004

Explorative approaches such as DNA microarray experiments are becoming increasingly important in microbial research. Despite these major technical advancements, approaches to study multifactor experiments are still lacking. We have addressed this problem by using rotation testing and a novel multivariate analysis of variance (MANOVA) approach (50-50 MANOVA) to investigate interacting experimental factors in a complex experimental design. Furthermore, a new rotation testing based method was introduced to calculate false-discovery rates for each response. This novel analytical concept was used to investigate global survival mechanisms in the environment of the major food-borne pathogen C. jejuni. We simulated nongrowth environmental conditions by investigating combinations of the factors temperature (5 and 25°C) and oxygen tension (anaerobic, microaerobic, and aerobic). Data were generated with DNA microarrays for information about gene expression patterns and Fourier transform infrared (FT-IR) spectroscopy to study global macromolecular changes in the cell. Microarray analyses showed that most genes were either unchanged or down regulated compared to the reference (day 0) for the conditions tested and that the 25°C anaerobic condition gave the most distinct expression pattern with the fewest genes expressed. The few up-regulated genes were generally stress related and/or related to the cell envelope. We found, using FT-IR spectroscopy, that the amount of polysaccharides and oligosaccharides increased under the nongrowth survival conditions. Potential mechanisms for survival could be to down regulate most functions to save energy and to produce polysaccharides and oligosaccharides for protection against harsh environments. Basic knowledge about the survival mechanisms is of fundamental importance in preventing transmission of this bacterium through the food chain.

* Corresponding author: Mailing address: Matforsk, Norwegian Food Research Institute, Osloveien 1, N-1430 Ås, Norway. Phone: 47 64 97 01 00. Fax: 47 64 97 03 33. E-mail for Birgitte Moen: birgitte.moen{at}matforsk.no. E-mail for Knut Rudi: knut.rudi{at}matforsk.no.

Present address: Department of Genetics, University of Leicester, Leicester LE1 7RH, United Kingdom.

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