Comparative transcriptome and phenotype analysis of acid-stressed Bacillus cereus strain ATCC 14579
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ABSTRACT:
ORGANISM(S): Bacillus cereus ATCC 14579 Bacillus cereus
PROVIDER: GSE13711 | GEO | 2009/11/25
SECONDARY ACCESSION(S): PRJNA114641
REPOSITORIES: GEO
Similar Datasets
Project description:The food-borne human pathogen Bacillus cereus is found in environments that often have a low pH, such as food and soil. The physiological response upon exposure to several levels of acidity were investigated of B. cereus model strain ATCC 10987, to elucidate the response of B. cereus to acid stress. pH 5.4, pH 5.0, pH 4.8 and pH 4.5 were selected to conduct microarray analyses, based on the differences in physiological response upon exposure to the acid conditions. The transcriptome data revealed response specific profiles. Showing mechanisms induced upon all the different acid down-shocks, such as nitrate reductase and energy production genes, and several genes specifically expressed differentially in mild or lethal levels of acidity, such as F1F0-ATPase and cydAB. Furthermore, mechanisms involved in oxidative stress response were found highly up-regulated in response to both mild and lethal acid stress. The induction of oxidative stress related genes may be a response to the formation of reactive oxygen species by a perturbation of the electron transport chain. Therefore, the formation of hydroxyl radicals and/ or peroxynitrite was monitored upon exposure to the different levels of acidity with a fluorescent probe in a flow cytometer. The formation of these oxidative compounds was shown to be specific for lethal pHs and a model to relate radical formation with the observed transcriptome profiles was proposed.
2009-11-25 | GSE13729 | GEO
Project description:The food-borne human pathogen Bacillus cereus is found in environments that often have a low pH, such as food and soil. The physiological response upon exposure to several levels of acidity were investigated of B. cereus model strain ATCC 14579, to elucidate the response of B. cereus to acid stress. pH 5.4, pH 5.0, pH 4.8 and pH 4.5 were selected to conduct microarray analyses, based on the differences in physiological response upon exposure to the acid conditions. The transcriptome data revealed response specific profiles. Showing mechanisms induced upon all the different acid down-shocks, such as nitrate reductase and energy production genes, and several genes specifically expressed differentially in mild or lethal levels of acidity, such as F1F0-ATPase and cydAB. Furthermore, mechanisms involved in oxidative stress response were found highly up-regulated in response to both mild and lethal acid stress. The induction of oxidative stress related genes may be a response to the formation of reactive oxygen species by a perturbation of the electron transport chain. Therefore, the formation of hydroxyl radicals and/ or peroxynitrite was monitored upon exposure to the different levels of acidity with a fluorescent probe in a flow cytometer. The formation of these oxidative compounds was shown to be specific for lethal pHs and a model to relate radical formation with the observed transcriptome profiles was proposed. Per acid down-shock three exposure times (i.e., 10, 30 and 60 min) were each compared with non-exposed cells (i.e., t0). In total 4 different acid down-shocks were applied, pH 5.4, pH 5.0, pH 4.8 and pH 4.5. The experiments were performed in duplicate and the duplicate samples were hybridised with a dye-swap.
2010-06-25 | E-GEOD-13711 | biostudies-arrayexpress
Project description:The food-borne human pathogen Bacillus cereus is found in environments that often have a low pH, such as food and soil. The physiological response upon exposure to several levels of acidity were investigated of B. cereus model strain ATCC 10987, to elucidate the response of B. cereus to acid stress. pH 5.4, pH 5.0, pH 4.8 and pH 4.5 were selected to conduct microarray analyses, based on the differences in physiological response upon exposure to the acid conditions. The transcriptome data revealed response specific profiles. Showing mechanisms induced upon all the different acid down-shocks, such as nitrate reductase and energy production genes, and several genes specifically expressed differentially in mild or lethal levels of acidity, such as F1F0-ATPase and cydAB. Furthermore, mechanisms involved in oxidative stress response were found highly up-regulated in response to both mild and lethal acid stress. The induction of oxidative stress related genes may be a response to the formation of reactive oxygen species by a perturbation of the electron transport chain. Therefore, the formation of hydroxyl radicals and/ or peroxynitrite was monitored upon exposure to the different levels of acidity with a fluorescent probe in a flow cytometer. The formation of these oxidative compounds was shown to be specific for lethal pHs and a model to relate radical formation with the observed transcriptome profiles was proposed. Per acid down-shock three exposure times (i.e., 10, 30 and 60 min) were each compared with non-exposed cells (i.e., t0). In total 4 different acid down-shocks were applied, pH 5.4, pH 5.0, pH 4.8 and pH 4.5. The experiments were performed in duplicate and the duplicate samples were hybridised with a dye-swap.
2010-06-25 | E-GEOD-13729 | biostudies-arrayexpress
Project description:Comparative phenotype and transcriptome analyses were performed with Bacillus cereus ATCC 14579 exposed to acid down-shock to pH 5.5 set with different acidulants. When acidified with hydrochloric acid (HCl), growth was diminished, whereas 2 mM undissociated lactic acid (HL) or acetic acid (HAc) stopped growth without inactivation (bacteriostatic condition), and 15 mM undissociated HAc caused growth arrest and, finally, cell death, as reflected by a 3 to 4 log inactivation (bactericidal condition). Within the first 60 min after pH down-shock, the intracellular ATP levels of cultures shocked with HCl were increased. The bacteriostatic pH shocks did not result in increased nor decreased intracellular ATP levels, indicating that the high energy status within the stressed aerobically grown B. cereus cells could be maintained. In contrast, exposure to 15 mM undissociated HAc resulted in significant lower ATP levels, which was in accordance with the observed inactivation. The transcriptomic responses pH down-shocked cultures were studied in the same time frame. The analyses revealed general and specific responses coupled to the different phenotypes and the acidulant used. The general acid stress response, shown in all different pH shocks, involves modulation of pyruvate metabolism and an oxidative stress response. The shifts in pyruvate metabolism include induction dehydrogenases of a butanediol fermentation pathway under non-lethal acid stress conditions and of lactate, formate, and ethanol fermentation pathways under 15 mM HAc stress. Other 15 mM HAc-specific responses were induction of the alternative electron-transport systems, including cydAB, and fatty acid biosynthesis genes. Differences in gene expression for the bacteriostatic organic acid stress conditions compared to the growth-retarded inorganic stress condition indicated a more stringent oxidative stress response, including induction of an additional catalase gene and a gene encoding a Dps-like protein. Moreover, modulations in amino acid and oligopeptide transport were also found for the 2 mM HAc and HL shocks. HL-specific and HAc-specific responses both involve amino acid metabolism. Our study on the genome-wide responses of aerobically grown B. cereus pH 5.5 shocks provides a unique overview of the different responses induced by three acidulants relevant for food preservation.
2009-09-30 | GSE15140 | GEO
Project description:Comparative phenotype and transcriptome analyses were performed with Bacillus cereus ATCC 14579 exposed to acid down-shock to pH 5.5 set with different acidulants. When acidified with hydrochloric acid (HCl), growth was diminished, whereas 2 mM undissociated lactic acid (HL) or acetic acid (HAc) stopped growth without inactivation (bacteriostatic condition), and 15 mM undissociated HAc caused growth arrest and, finally, cell death, as reflected by a 3 to 4 log inactivation (bactericidal condition). Within the first 60 min after pH down-shock, the intracellular ATP levels of cultures shocked with HCl were increased. The bacteriostatic pH shocks did not result in increased nor decreased intracellular ATP levels, indicating that the high energy status within the stressed aerobically grown B. cereus cells could be maintained. In contrast, exposure to 15 mM undissociated HAc resulted in significant lower ATP levels, which was in accordance with the observed inactivation. The transcriptomic responses pH down-shocked cultures were studied in the same time frame. The analyses revealed general and specific responses coupled to the different phenotypes and the acidulant used. The general acid stress response, shown in all different pH shocks, involves modulation of pyruvate metabolism and an oxidative stress response. The shifts in pyruvate metabolism include induction dehydrogenases of a butanediol fermentation pathway under non-lethal acid stress conditions and of lactate, formate, and ethanol fermentation pathways under 15 mM HAc stress. Other 15 mM HAc-specific responses were induction of the alternative electron-transport systems, including cydAB, and fatty acid biosynthesis genes. Differences in gene expression for the bacteriostatic organic acid stress conditions compared to the growth-retarded inorganic stress condition indicated a more stringent oxidative stress response, including induction of an additional catalase gene and a gene encoding a Dps-like protein. Moreover, modulations in amino acid and oligopeptide transport were also found for the 2 mM HAc and HL shocks. HL-specific and HAc-specific responses both involve amino acid metabolism. Our study on the genome-wide responses of aerobically grown B. cereus pH 5.5 shocks provides a unique overview of the different responses induced by three acidulants relevant for food preservation. Per acid down-shock three exposure times (i.e., 10, 30 and 60 min) were each compared with non-exposed cells (i.e., t0). In total 4 different pH 5.5 acid down-shocks were applied. pH 5.5 was reached by adding different acidulants i.e., hydrochloric acid (HCl), lactic acid (HL) resulting in 2 mM undissociated HL, acetic acid (HAc) resulting in 15 mM undissociated HAc, and a combination of acetic acid and hydrochloric acid (HAc/HCl) resulting in 2 mM undissociated HAc. The experiments were performed in duplicate and the duplicate samples were hybridised with a dye-swap.
2010-06-25 | E-GEOD-15140 | biostudies-arrayexpress
Project description:In order to cause disease, the food- and water-borne pathogen Campylobacter jejuni must face the extreme acidity of the host stomach as well as cope with pH fluctuations in the intestine. In the present study, C. jejuni NCTC 11168 was grown under mild acidic conditions mimicking those encountered in the intestine. The resulting transcriptional profiles revealed how this bacterium fine-tunes gene expression in response to acid stress. This adaptation involves differential expression of respiratory pathways, induction of genes for phosphate transport and repression of energy generation and intermediary metabolism genes. Keywords: acid shock; dose response; transcriptional response to 3 mild-acidic pH growth conditions (pH6.5, pH 6.0 and pH5.0)
2008-01-21 | GSE9920 | GEO
Project description:While significant advances have been made in EHEC pathogenesis, we still do not fully understand the impact of environmental stress on EHEC virulence. During the course of infection, EHEC must evade or overcome several biological barriers, the first of which is the gastric acidity encountered during passage through the stomach. EHEC is remarkable in its ability to tolerate this acidity. There are four different acid resistance systems that provide E. coli O157:H7 protection against exposure to low pH (2-2.5). Interestingly, EHEC uses these acid resistance systems differentially for survival in foods versus the bovine intestinal tract. The glutamate-dependent acid-resistance system is thought to offer the best protection below pH 3. Since the infectious dose of EHEC is so low (50-100 organisms), acid resistance becomes an important virulence trait. Studies of EHEC response to acid stress have focused primarily on levels of acid tolerance and the molecular basis of tolerance. However, the impact of acid stress on EHEC virulence is less well understood. In the related pathogen, EPEC, the plasmid-encoded regulator, Per, that regulates expression of many EPEC virulence factors, is regulated negatively at pH 5.5 and positively at pH 8.0, suggesting that virulence gene expression is repressed during mild acid stress and enhanced in alkaline pH typical of the small intestine. Expression of EPEC type III secreted factors involved in A/E lesion formation has been shown to be influenced by factors including culture media, iron and calcium levels. Protein secretion was inhibited at pH 6 and 8. In a third study, a gadE (encoding acid resistance regulator) mutation resulted in increased adhesion of E.coli O157:H7 to colonic epithelial cells, suggesting negative regulation of one or more adhesins. Other studies have reported that shiga toxin production is sensitive to culture conditions including pH. However, there are no studies of EHEC virulence changes after more severe acid stress nor studies linking stressed EHEC virulence phenotype with transcriptional changes. The goal of this study was to determine how acid stress affects EHEC virulence properties and through microarray analysis, define the genetic basis for these changes. Understanding how acid stress modulates the virulence potential of this pathogen is essential for delineating the pathogenesis of disease caused by EHEC infection and may offer novel approaches to prevent and treat EHEC infections. Bacteria were grown in LB broth overnight, then subcultured into DMEM and grown at 37C, 5%Co2. Bacteria were then subjected to one of three acid stress protocols: 1) UA30: growth in DMEM pH 7.4 followed by growth in DMEM pH 3.0 for 30 minutes; 2) AA30: growth in DMEM pH 5.0 (adaptation) followed by growth in DMEM pH 3.0; 3) UA15: growth in DMEM pH 7.4 followed by growth in DMEM pH 3.0 for 15 minutes. DMEM was supplemented with 25 mM MES (pH 5.0) and in the case of the control (unadapted, unshocked) 25 mM MOPS (pH 7.4) and the adaptation step was again carried out at 37C and 5% CO2. Acid shocking was done at pH 3.0 (unbuffered) at room temperature for all treatments
2010-05-16 | E-GEOD-14069 | biostudies-arrayexpress
Project description:While significant advances have been made in EHEC pathogenesis, we still do not fully understand the impact of environmental stress on EHEC virulence. During the course of infection, EHEC must evade or overcome several biological barriers, the first of which is the gastric acidity encountered during passage through the stomach. EHEC is remarkable in its ability to tolerate this acidity. There are four different acid resistance systems that provide E. coli O157:H7 protection against exposure to low pH (2-2.5). Interestingly, EHEC uses these acid resistance systems differentially for survival in foods versus the bovine intestinal tract. The glutamate-dependent acid-resistance system is thought to offer the best protection below pH 3. Since the infectious dose of EHEC is so low (50-100 organisms), acid resistance becomes an important virulence trait. Studies of EHEC response to acid stress have focused primarily on levels of acid tolerance and the molecular basis of tolerance. However, the impact of acid stress on EHEC virulence is less well understood. In the related pathogen, EPEC, the plasmid-encoded regulator, Per, that regulates expression of many EPEC virulence factors, is regulated negatively at pH 5.5 and positively at pH 8.0, suggesting that virulence gene expression is repressed during mild acid stress and enhanced in alkaline pH typical of the small intestine. Expression of EPEC type III secreted factors involved in A/E lesion formation has been shown to be influenced by factors including culture media, iron and calcium levels. Protein secretion was inhibited at pH 6 and 8. In a third study, a gadE (encoding acid resistance regulator) mutation resulted in increased adhesion of E.coli O157:H7 to colonic epithelial cells, suggesting negative regulation of one or more adhesins. Other studies have reported that shiga toxin production is sensitive to culture conditions including pH. However, there are no studies of EHEC virulence changes after more severe acid stress nor studies linking stressed EHEC virulence phenotype with transcriptional changes. The goal of this study was to determine how acid stress affects EHEC virulence properties and through microarray analysis, define the genetic basis for these changes. Understanding how acid stress modulates the virulence potential of this pathogen is essential for delineating the pathogenesis of disease caused by EHEC infection and may offer novel approaches to prevent and treat EHEC infections.
2009-06-26 | GSE14069 | GEO
Project description:Abiotic stresses disturb and limit nitrogen-fixing symbioses between rhizobia and their host legumes. In particular, the effect of extracellular acidity on rhizobia has been taken as a model example for analysis because of the economic impact and worldwide distribution of these symbionts in agricultural countries. Except for valuable molecular-biological studies on different rhizobia, no consolidated models have been formulated to describe the central physiologic changes that occur in acid-stressed bacteria. We present here an integrated analysis entailing the main cultural, metabolic, and molecular responses of the model bacterium Sinorhizobium meliloti growing under controlled acid stress in a chemostat. A stepwise extracellular acidification of the culture medium had indicated that S. meliloti stopped growing at ca. pH 6.0â??6.1. Under such a limiting acid stress the rhizobia increased the O2 consumption per cell by more than 5-fold. This phenotype, together with an increase in the transcripts for several membrane cytochromes, entails a remarkably higher aerobic-respiration rate in acid-stressed rhizobia. Changes in the transcripts encoding enzymes for lipid biosynthesis were also observed, consistent with previous data on rhizobial pH-dependent membrane remodelling. Together with increased energy demands under acidity, proteomic and transcriptomic data revealed that while at pH 7.0 the transport and biosynthesis of cellular compounds were quite active processes, under acid stress most overexpressed markers were associated with protein biosynthesis, macromolecular degradation and/or recycling, and energy metabolism. Within this context, the pentose-phosphate pathway exhibited increases in several transcripts, enzymes, and metabolites. Moreover, multivariate analysis of global metabolome data (more than 60 compounds) served to unequivocally correlate the specific-metabolite profiles with the extracellular pH for growth, with strikingly sensitive variations being observed in the rhizobial metabolomes upon extracellular-pH changes of less than 0.5 units. Except for a ca. 120-kb DNA stretch within the pSymA no specific genomic regions were associated with the observed acid-stress responses. Further practical analyses should be focussed on the phenotypic impact and time course of the observed changes during the acid-stress perception and on the search for common responses during the previously described sublethal acid-adaptive processes in rhizobia pH 6.1 vs pH 7
2015-10-29 | E-GEOD-74449 | biostudies-arrayexpress
Project description:Antimicrobial chemicals are widely applied to clean and disinfect food-contacting surfaces. However, the cellular response of bacteria, such as Bacillus cereus, to various disinfectants is unclear. In this study, the physiological and genome-wide transcriptional responses of B. cereus ATCC 14579 exposed to four different disinfectants (i.e., benzalkonium chloride, sodium hypochlorite, hydrogen peroxide, and peracetic acid) were analyzed. The physiological response of B. cereus to different concentrations of the disinfectants used was investigated. For each disinfectant, concentrations leading to the attenuation of growth, growth arrest, and cell death were studied in more detail. The simultaneous analysis of the transcriptional responses of B. cereus upon exposure to the different concentrations of disinfectants revealed common responses induced by the four disinfectants. Notably, genes involved in the general and oxidative stress responses were commonly up-regulated. Furthermore, the obtained results indicate that all the disinfectants also induce specific responses. Exposure to benzalkonium chloride, a disinfectant known to induce membrane damage, specifically induced genes involved in the fatty acid metabolism. Benzalkonium chloride induced-membrane damage was confirmed by fluorescence microscopy and fatty acid analysis confirmed that fatty acid composition of cell membrane was affected upon exposure to benzalkonium chloride. Sodium hypochlorite induced genes involved in sulfur and sulfur-containing amino acids metabolism, which correlated with the observed sodium hypochlorite-specific induction of oxidation of sulphydryl groups. Hydrogen peroxide and peracetic acid exposures induced genes involved in DNA damage and the SOS response. Notably, hydrogen peroxide and peracetic acid-treated cells exhibited higher mutation rates corroborating with the induced SOS response. Understanding the mechanisms displayed by microorganisms coping with disinfectants-induced stress may allow for design of more efficient sequential and/or disinfectant combination treatments in food processing environments.
2010-03-31 | GSE18807 | GEO