An integrative approach to identify Bacillus weihenstephanensis resistance biomarkers using gene expression quantification throughout acid inactivation

被引:10
作者
Desriac, Noemie [1 ,2 ]
Coroller, Louis [2 ]
Sohier, Daniele [1 ]
Postollec, Florence [1 ]
机构
[1] ADRIA Dev, ZA Creach Gwen, UMT PHYSI Opt 08 3, F-29196 Quimper, France
[2] Univ Brest, Lab Univ Biodivers & Ecol Microbienne, EA3882, UMT PHYSI Opt 08 3,ScInBioS IFR148, F-29334 Quimper, France
关键词
Bacillus weihenstephanensis; Acid resistance; Gene expression; Modelling; Biomarker; Food safety; REVERSE TRANSCRIPTION-PCR; REAL-TIME PCR; LISTERIA-MONOCYTOGENES; VEGETATIVE CELLS; RISK-ASSESSMENT; CEREUS; STRESS; SURVIVAL; BACTERIA; ENVIRONMENTS;
D O I
10.1016/j.fm.2012.05.008
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The aim of this study was to define an integrative approach to identify resistance biomarkers using gene expression quantification and mathematical modelling. Mid-exponentially growing cells were transferred into acid conditions (BHI, pH 4.6) to obtain inactivation kinetics, performed in triplicate. The inactivation curve was fitted with a mixed Weibull model. This model allowed to differentiate two subpopulations with various acid resistances among the initial population. In parallel, differential gene expression was quantified by RT-qPCR. While narL was down-regulated throughout acid inactivation, sigB and katA were up-regulated. sigB expression up-regulation peak was correlated to the less resistant subpopulation when katA up-regulation, was correlated to the more resistant subpopulation. Moreover, differences in population structure were highlighted between each replicate. The higher proportion of the more resistant subpopulation was linked to a higher katA gene expression. These results suggest that sigB and katA might be used as different types of biomarkers, for instance to track moderate and high acid-resistance, respectively. The use of this approach combining RT-qPCR and predictive modelling to track cellular biomarker variations appears as an interesting tool to take into account physiological cell responses into mathematical modelling, allowing an accurate prediction of microbial behaviour. (C) 2012 Elsevier Ltd. All rights reserved.
引用
收藏
页码:172 / 178
页数:7
相关论文
共 47 条
[1]   Biomarkers and surrogate endpoints: Preferred definitions and conceptual framework [J].
Atkinson, AJ ;
Colburn, WA ;
DeGruttola, VG ;
DeMets, DL ;
Downing, GJ ;
Hoth, DF ;
Oates, JA ;
Peck, CC ;
Schooley, RT ;
Spilker, BA ;
Woodcock, J ;
Zeger, SL .
CLINICAL PHARMACOLOGY & THERAPEUTICS, 2001, 69 (03) :89-95
[2]   The wet-heat resistance of Bacillus weihenstephanensis KBAB4 spores produced in a two-step sporulation process depends on sporulation temperature but not on previous cell history [J].
Baril, E. ;
Coroller, L. ;
Postollec, F. ;
Leguerinel, I. ;
Boulais, C. ;
Carlin, F. ;
Mafart, P. .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 2011, 146 (01) :57-62
[3]   NONTHERMAL INACTIVATION MODELS FOR LISTERIA-MONOCYTOGENES [J].
BUCHANAN, RL ;
GOLDEN, MH ;
WHITING, RC ;
PHILLIPS, JG ;
SMITH, JL .
JOURNAL OF FOOD SCIENCE, 1994, 59 (01) :179-188
[4]   The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments [J].
Bustin, Stephen A. ;
Benes, Vladimir ;
Garson, Jeremy A. ;
Hellemans, Jan ;
Huggett, Jim ;
Kubista, Mikael ;
Mueller, Reinhold ;
Nolan, Tania ;
Pfaffl, Michael W. ;
Shipley, Gregory L. ;
Vandesompele, Jo ;
Wittwer, Carl T. .
CLINICAL CHEMISTRY, 2009, 55 (04) :611-622
[5]   Escherichia coli O157:H7 stress and virulence gene expression on Romaine lettuce using comparative real-time PCR [J].
Carey, Christine M. ;
Kostrzynska, Magdalena ;
Thompson, Stacey .
JOURNAL OF MICROBIOLOGICAL METHODS, 2009, 77 (02) :235-242
[6]   Comparative Transcriptomic and Phenotypic Analysis of the Responses of Bacillus cereus to Various Disinfectant Treatments [J].
Ceragioli, Mara ;
Mols, Maarten ;
Moezelaar, Roy ;
Ghelardi, Emilia ;
Senesi, Sonia ;
Abee, Tjakko .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2010, 76 (10) :3352-3360
[7]   Survival of Bacillus cereus spores and vegetative cells in acid media simulating human stomach [J].
Clavel, T ;
Carlin, F ;
Lairon, D ;
Nguyen-The, C ;
Schmitt, P .
JOURNAL OF APPLIED MICROBIOLOGY, 2004, 97 (01) :214-219
[8]   General model, based on two mixed Weibull distributions of bacterial resistance, for describing various shapes of inactivation curves [J].
Coroller, L. ;
Leguerinel, I. ;
Mettler, E. ;
Savy, N. ;
Mafart, P. .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (10) :6493-6502
[9]   A novel hybrid kinase is essential for regulating the σB-mediated stress response of Bacillus cereus [J].
de Been, Mark ;
Tempelaars, Marcel H. ;
van Schaik, Willem ;
Moezelaar, Roy ;
Siezen, Roland J. ;
Abee, Tjakko .
ENVIRONMENTAL MICROBIOLOGY, 2010, 12 (03) :730-744
[10]   Quantification of the effects of salt stress and physiological state on thermotolerance of Bacillus cereus ATCC 10987 and ATCC 14579 [J].
den Besten, Heidy M. W. ;
Mataragas, Marios ;
Moezelaar, Roy ;
Abee, Tjakko ;
Zwietering, Marcel H. .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2006, 72 (09) :5884-5894