scholarly journals Acid tolerance in early colonizers of oral biofilms

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Gabriella Boisen ◽  
Julia R. Davies ◽  
Jessica Neilands
Keyword(s):  
Acid Tolerance ◽  
Low Ph ◽  
Salivary Proteins ◽  
Confocal Scanning Laser Microscopy ◽  
Planktonic Cells ◽  
Tolerance Development ◽  
Salivary Pellicle ◽  
Oral Biofilms ◽  
Acid Tolerant

Abstract Background In caries, low pH drives selection and enrichment of acidogenic and aciduric bacteria in oral biofilms, and development of acid tolerance in early colonizers is thought to play a key role in this shift. Since previous studies have focussed on planktonic cells, the effect of biofilm growth as well as the role of a salivary pellicle on this process is largely unknown. We explored acid tolerance and acid tolerance response (ATR) induction in biofilm cells of both clinical and laboratory strains of three oral streptococcal species (Streptococcus gordonii, Streptococcus oralis and Streptococcus mutans) as well as two oral species of Actinomyces (A. naeslundii and A. odontolyticus) and examined the role of salivary proteins in acid tolerance development. Methods Biofilms were formed on surfaces in Ibidi® mini flow cells with or without a coating of salivary proteins and acid tolerance assessed by exposing them to a challenge known to kill non-acid tolerant cells (pH 3.5 for 30 min) followed by staining with LIVE/DEAD BacLight and confocal scanning laser microscopy. The ability to induce an ATR was assessed by exposing the biofilms to an adaptation pH (pH 5.5) for 2 hours prior to the low pH challenge. Results Biofilm formation significantly increased acid tolerance in all the clinical streptococcal strains (P < 0.05) whereas the laboratory strains varied in their response. In biofilms, S. oralis was much more acid tolerant than S. gordonii or S. mutans. A. naeslundii showed a significant increase in acid tolerance in biofilms compared to planktonic cells (P < 0.001) which was not seen for A. odontolyticus. All strains except S. oralis induced an ATR after pre-exposure to pH 5.5 (P < 0.05). The presence of a salivary pellicle enhanced both acid tolerance development and ATR induction in S. gordonii biofilms (P < 0.05) but did not affect the other bacteria to the same extent. Conclusions These findings suggest that factors such as surface contact, the presence of a salivary pellicle and sensing of environmental pH can contribute to the development of high levels of acid tolerance amongst early colonizers in oral biofilms which may be important in the initiation of caries.

Stress-responsive proteins are upregulated in Streptococcus mutans during acid tolerance

Microbiology ◽  
2004 ◽  
Vol 150 (5) ◽  
pp. 1339-1351 ◽  
Author(s):  
Alice C. L. Len ◽  
Derek W. S. Harty ◽  
Nicholas A. Jacques
Keyword(s):  
Streptococcus Mutans ◽  
Elongation Factor ◽  
Acid Tolerance ◽  
Proteome Analysis ◽  
Low Ph ◽  
Lactobacillus Helveticus ◽  
Oral Streptococci ◽  
Flight Mass Spectrometry ◽  
Future Assessment ◽  
Acid Tolerant

Streptococcus mutans is an important pathogen in the initiation of dental caries as the bacterium remains metabolically active when the environment becomes acidic. The mechanisms underlying this ability to survive and proliferate at low pH remain an area of intense investigation. Differential two-dimensional electrophoretic proteome analysis of S. mutans grown at steady state in continuous culture at pH 7·0 or pH 5·0 enabled the resolution of 199 cellular and extracellular protein spots with altered levels of expression. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry identified 167 of these protein spots. Sixty-one were associated with stress-responsive pathways involved in DNA replication, transcription, translation, protein folding and proteolysis. The 61 protein spots represented isoforms or cleavage products of 30 different proteins, of which 25 were either upregulated or uniquely expressed during acid-tolerant growth at pH 5·0. Among the unique and upregulated proteins were five that have not been previously identified as being associated with acid tolerance in S. mutans and/or which have not been studied in any detail in oral streptococci. These were the single-stranded DNA-binding protein, Ssb, the transcription elongation factor, GreA, the RNA exonuclease, polyribonucleotide nucleotidyltransferase (PnpA), and two proteinases, the ATP-binding subunit, ClpL, of the Clp family of proteinases and a proteinase encoded by the pep gene family with properties similar to the dipeptidase, PepD, of Lactobacillus helveticus. The identification of these and other differentially expressed proteins associated with an acid-tolerant-growth phenotype provides new information on targets for mutagenic studies that will allow the future assessment of their physiological significance in the survival and proliferation of S. mutans in low pH environments.

Augmentation of Killing of Escherichia coli O157 by Combinations of Lactate, Ethanol, and Low-pH Conditions

1999 ◽  
Vol 65 (3) ◽  
pp. 1308-1311 ◽  
Author(s):  
Sarah L. Jordan ◽  
Jayne Glover ◽  
Laura Malcolm ◽  
Fiona M. Thomson-Carter ◽  
Ian R. Booth ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Acid Tolerance ◽  
Low Ph ◽  
Exponential Phase ◽  
Escherichia Coli O157 ◽  
Cytoplasmic Ph ◽  
Ph Homeostasis ◽  
Acid Tolerant ◽  
Ph Conditions

ABSTRACT The acid tolerance of Escherichia coli O157:H7 strains can be overcome by addition of lactate, ethanol, or a combination of the two agents. Killing can be increased by as much as 4 log units in the first 5 min of incubation at pH 3 even for the most acid-tolerant isolates. Exponential-phase, habituated, and stationary-phase cells are all sensitive to incubation with lactate and ethanol. Killing correlates with disruption of the capacity for pH homeostasis. Habituated and stationary-phase cells can partially offset the effects of the lowering of cytoplasmic pH.

scholarly journals Genetic and Physiologic Analysis of thegroE Operon and Role of the HrcA Repressor in Stress Gene Regulation and Acid Tolerance in Streptococcus mutans

2001 ◽  
Vol 183 (20) ◽  
pp. 6074-6084 ◽  
Author(s):  
José A. C. Lemos ◽  
Yi-Ywan M. Chen ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Acid Tolerance ◽  
Oral Bacteria ◽  
Mrna Levels ◽  
Acid Shock ◽  
Oral Biofilms ◽  
In The Wild ◽  
Dnak Protein ◽  
Stress Gene

ABSTRACT Our working hypothesis is that the major molecular chaperones DnaK and GroE play central roles in the ability of oral bacteria to cope with the rapid and frequent stresses encountered in oral biofilms, such as acidification and nutrient limitation. Previously, our laboratory partially characterized the dnaK operon ofStreptococcus mutans(hrcA-grpE-dnaK) and demonstrated that dnaK is up-regulated in response to acid shock and sustained acidification (G. C. Jayaraman, J. E. Penders, and R. A. Burne, Mol. Microbiol. 25:329–341, 1997). Here, we show that thegroESL genes of S. mutans constitute an operon that is expressed from a stress-inducible ςA-type promoter located immediately upstream of a CIRCE element. GroEL protein and mRNA levels were elevated in cells exposed to a variety of stresses, including acid shock. A nonpolar insertion into hrcA was created and used to demonstrate that HrcA negatively regulates the expression of thegroEL and dnaK operons. The SM11 mutant, which had constitutively high levels of GroESL and roughly 50% of the DnaK protein found in the wild-type strain, was more sensitive to acid killing and could not lower the pH as effectively as the parent. The acid-sensitive phenotype of SM11 was, at least in part, attributable to lower F1F0-ATPase activity. A minimum of 10 proteins, in addition to GroES-EL, were found to be up-regulated in SM11. The data clearly indicate that HrcA plays a key role in the regulation of chaperone expression in S. mutans and that changes in the levels of the chaperones profoundly influence acid tolerance.

scholarly journals Nitrification in a Biofilm at Low pH Values: Role of In Situ Microenvironments and Acid Tolerance

2006 ◽  
Vol 72 (6) ◽  
pp. 4283-4292 ◽  
Author(s):  
Armin Gieseke ◽  
Sheldon Tarre ◽  
Michal Green ◽  
Dirk de Beer
Keyword(s):  
Ammonia Oxidation ◽  
Acid Tolerance ◽  
Low Ph ◽  
Nitrifying Bacteria ◽  
Experimental Conditions ◽  
Mass Transfer Limitation ◽  
Highly Active ◽  
Acid Tolerant ◽  
Acidic Conditions

ABSTRACT The sensitivity of nitrifying bacteria to acidic conditions is a well-known phenomenon and generally attributed to the lack and/or toxicity of substrates (NH3 and HNO2) with decreasing pHs. In contrast, we observed strong nitrification at a pH around 4 in biofilms grown on chalk particles and investigated the following hypotheses: the presence of less acidic microenvironments and/or the existence of acid-tolerant nitrifiers. Microelectrode measurements (in situ and under various experimental conditions) showed no evidence of a neutral microenvironment, either within the highly active biofilm colonizing the chalk surface or within a control biofilm grown on a nonbuffering (i.e., sintered glass) surface under acidic pH. A 16S rRNA approach (clone libraries and fluorescence in situ hybridizations) did not reveal uncommon nitrifying (potentially acid-tolerant) strains. Instead, we found a strongly acidic microenvironment, evidence for a clear adaptation to the low pH in situ, and the presence of nitrifying populations related to subgroups with low Km s for ammonia (Nitrosopira spp., Nitrosomonas oligotropha, and Nitrospira spp.). Acid-consuming (chalk dissolution) and acid-producing (ammonia oxidation) processes are equilibrated on a low-pH steady state that is controlled by mass transfer limitation through the biofilm. Strong affinity to ammonia and possibly the expression of additional functions, e.g., ammonium transporters, are adaptations that allow nitrifiers to cope with acidic conditions in biofilms and other habitats.

scholarly journals Acid Tolerance of Biofilm Cells of Streptococcus mutans

2007 ◽  
Vol 73 (17) ◽  
pp. 5633-5638 ◽  
Author(s):  
Jessica Welin-Neilands ◽  
Gunnel Svensäter
Keyword(s):  
Streptococcus Mutans ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Planktonic Cells ◽  
Viable Cells ◽  
Tolerance Response ◽  
Acid Tolerant ◽  
Carious Process ◽  
Different Strains ◽  
Viability Stain

ABSTRACT Streptococcus mutans, a member of the dental plaque community, has been shown to be involved in the carious process. Cells of S. mutans induce an acid tolerance response (ATR) when exposed to sublethal pH values that enhances their survival at a lower pH. Mature biofilm cells are more resistant to acid stress than planktonic cells. We were interested in studying the acid tolerance and ATR-inducing ability of newly adhered biofilm cells of S. mutans. All experiments were carried out using flow-cell systems, with acid tolerance tested by exposing 3-h biofilm cells to pH 3.0 for 2 h and counting the number of survivors by plating on blood agar. Acid adaptability experiments were conducted by exposing biofilm cells to pH 5.5 for 3 h and then lowering the pH to 3.5 for 30 min. The viability of the cells was assessed by staining the cells with LIVE/DEAD BacLight viability stain. Three-hour biofilm cells of three different strains of S. mutans were between 820- and 70,000-fold more acid tolerant than corresponding planktonic cells. These strains also induced an ATR that enhanced the viability at pH 3.5. The presence of fluoride (0.5 M) inhibited the induction of an ATR, with 77% fewer viable cells at pH 3.5 as a consequence. Our data suggest that adhesion to a surface is an important step in the development of acid tolerance in biofilm cells and that different strains of S. mutans possess different degrees of acid tolerance and ability to induce an ATR.

scholarly journals Regulation and Physiological Significance of ClpC and ClpP in Streptococcus mutans

2002 ◽  
Vol 184 (22) ◽  
pp. 6357-6366 ◽  
Author(s):  
José A. C. Lemos ◽  
Robert A. Burne
Keyword(s):  
Gene Expression ◽  
Streptococcus Mutans ◽  
Stress Responses ◽  
Expression Patterns ◽  
Acid Tolerance ◽  
Low Ph ◽  
Two Dimensional ◽  
Wild Type ◽  
Protein Gels

ABSTRACT Tolerance of environmental stress, especially low pH, by Streptococcus mutans is central to the virulence of this organism. The Clp ATPases are implicated in the tolerance of, and regulation of the response to, stresses by virtue of their protein reactivation and remodeling activities and their capacity to target misfolded proteins for degradation by the ClpP peptidase. The purpose of this study was to dissect the role of selected clp genes in the stress responses of S. mutans, with a particular focus on acid tolerance and adaptation. Homologues of the clpB, clpC, clpE, clpL, clpX, and clpP genes were identified in the S. mutans genome. The expression of clpC and clpP, which were chosen as the focus of this study, was induced at low pH and at growth above 40°C. Inactivation of ctsR, the first of two genes in the clpC operon, demonstrated that CtsR acts as a repressor of clp and groES-EL gene expression. Strains lacking ClpP, but not strains lacking ClpC, were impaired in their ability to grow under stress-inducing conditions, formed long chains, aggregated in culture, had reduced genetic transformation efficiencies, and had a reduced capacity to form biofilms. Comparison of two-dimensional protein gels from wild-type cells and the ctsR and clpP mutants revealed many changes in the protein expression patterns. In particular, in the clpP mutant, there was an increased production of GroESL and DnaK, suggesting that cells were stressed, probably due to the accumulation of denatured proteins.

scholarly journals The role of resistance to bile salts and acid tolerance of exopolysaccharides (EPSS) produced by yogurt starter bacteria

2010 ◽  
Vol 62 (2) ◽  
pp. 323-328 ◽  
Author(s):  
Hatice Boke ◽  
Belma Aslim ◽  
Gulcin Alp
Keyword(s):  
Protective Effect ◽  
Bile Salts ◽  
Acid Tolerance ◽  
Streptococcus Thermophilus ◽  
Low Ph ◽  
Lactobacillus Delbrueckii ◽  
Viable Bacteria ◽  
Lactobacillus Delbrueckii Subsp

The aim of this study was to investigate a possible relation between EPS production and resistance to bile salts and tolerance to low pH. Eight strains which produced the highest and lowest amount of EPS (16- 211mg/l) were selected among 54 bacteria isolated from yogurt. Additionally, they were tested for resistance to bile salts (0.15, 0.3 %) and tolerance to low pH (2.0-3.0). After treatment with bile salts and acid, viable bacteria (log cfu ml-1) were determined by surface plating. The high EPS producing strains (B3, G12, W22) showed a significant (P<0.05) protective effect against low pH (pH 2.0). All Streptococcus thermophilus strains showed a higher tolerance to bile salts than the Lactobacillus delbrueckii subsp. bulgaricus strains. The high EPS-producing S. thermophilus (W22, T12) and L. bulgaricus (B3, G2) strains showed a significant (P<0.01) protective effect against bile salts (0.3 %).

scholarly journals Differences in Salivary Proteins as a Function of PROP Taster Status and Gender in Normal Weight and Obese Subjects

Molecules ◽  
2021 ◽  
Vol 26 (8) ◽  
pp. 2244
Author(s):  
Melania Melis ◽  
Mariano Mastinu ◽  
Stefano Pintus ◽  
Tiziana Cabras ◽  
Roberto Crnjar ◽  
...  
Keyword(s):  
Food Preferences ◽  
Specific Weight ◽  
Normal Weight ◽  
Nutrition Status ◽  
Salivary Proteins ◽  
Specific Proteins ◽  
Obese Subjects ◽  
And Gender ◽  
Cystatin Sn

Taste plays an important role in processes such as food choices, nutrition status and health. Salivary proteins contribute to taste sensitivity. Taste reduction has been associated with obesity. Gender influences the obesity predisposition and the genetic ability to perceive the bitterness of 6-n-propylthiouracil (PROP), oral marker for food preferences and consumption. We investigated variations in the profile of salivary proteome, analyzed by HPLC-ESI-MS, between sixty-one normal weight subjects (NW) and fifty-seven subjects with obesity (OB), based on gender and PROP sensitivity. Results showed variations of taste-related salivary proteins between NW and OB, which were differently associated with gender and PROP sensitivity. High levels of Ps-1, II-2 and IB-1 proteins belonging to basic proline rich proteins (bPRPs) and PRP-1 protein belonging to acid proline rich proteins (aPRPs) were found in OB males, who showed a lower body mass index (BMI) than OB females. High levels of Ps-1 protein and Cystatin SN (Cyst SN) were found in OB non-tasters, who had lower BMI than OB super-tasters. These new insights on the role of salivary proteins as a factor driving the specific weight gain of OB females and super-tasters, suggest the use of specific proteins as a strategic tool modifying taste responses related to eating behavior.

scholarly journals Role of Anions in Low pH-induced Translocation of Diphtheria Toxin

1989 ◽  
Vol 264 (19) ◽  
pp. 11367-11372 ◽  
Author(s):  
J Ø Moskaug ◽  
K Sandvig ◽  
S Olsnes
Keyword(s):  
Diphtheria Toxin ◽  
Low Ph
Sign in / Sign up

Export Citation Format

Share Document