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Study of Bacillus cereus spore germination and outgrowth in a vegetable food model by means of genome-wide transcriptome analysis

ABSTRACT: Pathogenic species belonging to Bacillus cereus sensu lato group possess a high evolutionary advantage in the environment and in food matrices thanks to their capacity to survive as silent spores to harsh environmental insults and grow at relatively low temperatures. Ready to re-heat products are at severe risk for contamination by members of Bacillus cereus s.l. group if not stored at proper conditions. In this work, the goal was to assess, by means of a genome-wide transcriptional assay, the isolated strain Bacillus cereus UC10070 gene expression behind the process of spore germination and consequent outgrowth in an artificially contaminated vegetable-based food model. A vegetable food model subjected to a heat treatment was determined to present favourable conditions for spores germination. Microscopic analyses together with OD measurements were applied to select the key steps of B. cereus cell cycle to be used for the microarray analysis. Using this approach we found a total of 1,646 probe sets differentially expressed and modulated during the entire B. cereus life cycle in the vegetable foodstuff.

ORGANISM(S): Bacillus cereus [Bacillus thuringiensis] serovar konkukian str. 97-27

PROVIDER: GSE65259 | GEO | 2015/01/26

SECONDARY ACCESSION(S): PRJNA273561

REPOSITORIES: GEO

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Study of Bacillus cereus spore germination and outgrowth in a vegetable food model by means of genome-wide transcriptome analysis

Project description:Pathogenic species belonging to Bacillus cereus sensu lato group possess a high evolutionary advantage in the environment and in food matrices thanks to their capacity to survive as silent spores to harsh environmental insults and grow at relatively low temperatures. Ready to re-heat products are at severe risk for contamination by members of Bacillus cereus s.l. group if not stored at proper conditions. In this work, the goal was to assess, by means of a genome-wide transcriptional assay, the isolated strain Bacillus cereus UC10070 gene expression behind the process of spore germination and consequent outgrowth in an artificially contaminated vegetable-based food model. A vegetable food model subjected to a heat treatment was determined to present favourable conditions for spores germination. Microscopic analyses together with OD measurements were applied to select the key steps of B. cereus cell cycle to be used for the microarray analysis. Using this approach we found a total of 1,646 probe sets differentially expressed and modulated during the entire B. cereus life cycle in the vegetable foodstuff. RNA was isolated from the dormant spores (DS), germinating spores at 40 minutes (GSP), early-log phase at 2h (C2h) and late log phase cells at 12 hours (C12h) after thermal treatment of B. cereus UC10070 previously isolated froma biofilm on a spoiled vegetable-based puree. There were 3 biological replicates (independent cultures) for each condition. Complete genome sequence of B. thuringiensis sv konkukian str. 97-27 (NCBI Reference Sequence: NC_005957.1) was chosen for its high homology with B. cereus s.l. UC10070, to design probes corresponding to 5,197 genes spotted in duplicates onto ElectraSenseH 12K microarrays chip.

2015-01-26 | E-GEOD-65259 | biostudies-arrayexpress

Identification of CdnL, a putative transcriptional regulator involved in repair and outgrowth of heat-damaged Bacillus cereus spores

Project description:Heat-treated spores show delayed and slower germination and outgrowth compared to untreated spores presumably due to spore damage repair. This study was performed to identify genes possibly involved in spore damage repair in B. cereus. In this study we compared the transcriptomic profiles of untreated and heat-treated spores during germination and outgrowth in BHI at 30C.

2016-02-02 | GSE73043 | GEO

Dynamics of Germinosome Formation Interactions between GerD and Germinant Receptor Subunits in Bacillus cereus Spores

Project description:Spores of the bacterium Bacillus cereus can cause disease in humans due to contamination of raw materials for food manufacturing. These dormant, resistant spores can survive for years in the environment, but can germinate and grow when their surroundings become suitable, and spore germination proteins play an important role in the decision to germinate. The dataset attached describes the quantification of the fluorophores of the expressed fusion proteins by label free mass spectrometry.

2022-01-24 | MSV000088726 | MassIVE

Temperature dependence of the proteome profile of the psychrotolerant pathogenic food spoiler Bacillus weihenstephanensis type strain WSBC 10204

Project description:Bacillus weihenstephanensis is a subspecies of the Bacillus cereus sensu lato group of spore forming bacteria known to cause food spoilage or food poisoning. The key distinguishing phenotype of B. weihenstephanensis is its ability to grow below 7°C or, from a food safety perspective, to grow and potentially produce toxins in a refrigerated environment. In order to gain insight into to the mechanistic basis of its psychrotolerant phenotype, as well as elucidate relevant aspects of its toxigenic profile, the proteome profiles of cells grown at either 6°C or 30°C were compared.

2015-12-01 | PXD002007 | Pride

Stoichiometry and absolute abundance of proteins in the Bacillus cereus spore insoluble fraction determined using a QconCAT approach

Project description:Spores of Bacillus cereus pose a threat to food safety due to their high resistance to the heat or acid treatments commonly used to make food microbially safe. As spores survive these treatments and later resume growth either on foodstuffs or, after ingestion, upon entering the gut they are capable of producing toxins which cause either vomiting or diarrhea respectively. The outer layers of the spore consist primarily of proteins which may serve as potential biomarkers for detection. In order to quantify proteins in the insoluble fraction of the spore coat, a proteomics approach using a QconCAT reference standard was employed which is the first time this is used in a heterogeneous system. This allowed us to determine the abundance of 21 proteins which spanned across three orders of magnitude and together covered 5.66% ±0.51 of the total spore weight. Classification by abundance resulted in functional characterisation of the protein BC_0987, renamed SasS. Furthermore, protein stoichiometry and determination of the abundance of, for instance, germination mediating enzymes provides useful information for model development. And finally, the four most abundant proteins, CotB1, CotB2, CotX and InhA, are presented to be biomarker candidates containing immunogenic epitopes as predicted by the SVMTriP algorithm and being conveniently located in the outermost layers of the spore.

2018-10-26 | PXD003569 | Pride

‘One-pot’ sample processing method for proteome-wide analysis of microbial cells and spores

Project description:Bacterial endospores, the transmissible forms of pathogenic bacilli and clostridia, are heterogeneous multilayered structures composed of proteins. These proteins protect the spores against variety of stresses, thus helping spore survival, and assist in germination, by interacting with the environment to form vegetative cells. Owing to the complexity, insolubility, and dynamic nature of spore proteins, it has been difficult to obtain their comprehensive protein profiles. The intact spores of Bacillus subtilis, Bacillus cereus, and Peptoclostridium difficile and their vegetative counterparts were disrupted by bead-beating in 6M urea under reductive conditions. The heterogeneous mixture was then double-digested with LysC and trypsin. Next, the peptide mixture was pre-fractionated with Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) followed by reverse phase LC-FT-MS analysis of the fractions. ‘One-pot’ method is a simple, robust method that yields identification of >1000 proteins with high confidence, across all spore layers from Bacillus subtilis, Bacillus cereus, and Peptoclostridium difficile. This method can be employed for proteome-wide analysis of non-spore-forming as well as spore-forming pathogens. Analysis of spore protein profile will help to understand the sporulation and germination processes and to distinguish immunogenic protein markers.

2018-07-05 | PXD008242 | Pride

Distinct roles of ComK1 and ComK2 in gene regulation in Bacillus cereus (overexpressed ComK)

Project description:Comparison of the Bacillus cereus with overexpressed Bacillus subtilis ComK (Bacillus cereus pNWcomKBsu) vs Bacillus cereus carrying empty plasmid (Bacillus cereus pNW33N)

2011-07-28 | GSE27265 | GEO

The sigmaB regulon of B. cereus ATCC14579

Project description:The alternative sigma factor sigmaB has an important role in the acquisition of stress-resistance in many gram-positive bacteria. In the foodborne pathogen Bacillus cereus sigmaB is activated strongly upon a heat shock and other stress conditions. Here we describe the identification of the set of sigmaB-regulated genes in B. cereus by DNA microarray analysis of the transcriptome upon a mild heat shock. A total of 24 genes could be identified as being sigmaB-dependent as witnessed by (i) significantly lower expression-levels of these genes in mutants deleted for sigB and rsbY (encoding the alternative sigma factor sigmaB and a PP2C phosphatase that acts as a crucial positive regulator of sigmaB-activity, respectively) than in the parental strain B. cereus ATCC 14579, and (ii) increased expression of these genes upon a heat shock. Newly identified members of the sigmaB-regulon of B. cereus include an ECF-sigma factor (sigmaZ), a histidine kinase and two genes that have predicted functions in spore germination. Our data indicate that the sigmaB-regulon of B. cereus is considerably smaller than that of other gram-positive bacteria. This appears to be in line with phylogenetic analyses of sigmaB in which sigmaB in the B. cereus group was suggested to be close to the ancestral form of sigmaB in gram-positive bacteria. Keywords: stress response, comparative genomics

2007-04-10 | GSE6005 | GEO

The sigmaB regulon of B. cereus ATCC14579

2010-07-01 | E-GEOD-6005 | biostudies-arrayexpress

Expression analysis of sporulating Bacillus subtilis strain 168 deleted for prpE gene

Project description:Investigation of whole genome gene expression level changes in sporulating Bacillus subtilis 168 delta-prpE mutant, compared to the wild-type strain. The mutation engineered into this strain results in impaired germination of spores.

2013-07-08 | GSE44125 | GEO

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