Bacterial Population Size Research Articles

Evolution of mutation rates in bacteria

Evolutionary success of bacteria relies on the constant fine-tuning of their mutation rates, which optimizes their adaptability to constantly changing environmental conditions. When adaptation is limited by the mutation supply rate, under some conditions, natural selection favours increased mutation rates by acting on allelic variation of the genetic systems that control fidelity of DNA replication and repair. Mutator alleles are carried to high frequency through hitchhiking with the adaptive mutations they generate. However, when fitness gain no longer counterbalances the fitness loss due to continuous generation of deleterious mutations, natural selection favours reduction of mutation rates. Selection and counter-selection of high mutation rates depends on many factors: the number of mutations required for adaptation, the strength of mutator alleles, bacterial population size, competition with other strains, migration, and spatial and temporal environmental heterogeneity. Such modulations of mutation rates may also play a role in the evolution of antibiotic resistance.

A review of issues related to the quantification of bacteria from the phyllosphere

Quantification of the size of epiphytic bacterial populations and characterization of their composition involves definition of a sampling strategy in time and space, the choice of methods for liberating the bacteria from the leaf surface and for recovering them for subsequent determination of the number of viable or culturable cells. This literature review focuses on some of the issues related to these choices. After briefly reviewing the different types of epiphytic colonizers we consider the biological, methodological, and statistical consequences of the choice of the sampling unit and of the spatial and temporal variability of population size and composition for epiphytic bacteria. The different methods available for the detection and enumeration of naturally occurring microorganisms in the phyllosphere are discussed. Advantages and drawbacks of each are described in this review designed as a ‘hands-on’ guide.

The Variation of Nitrifying Bacterial Population Sizes in a Sequencing Batch Reactor (SBR) Treating Low/Mid/High Concentrated Wastewater

The Variation of Nitrifying Bacterial Population Sizes in a Sequencing Batch Reactor (SBR) Treating Low/Mid/High Concentrated Wastewater

Modeling suggests frequency estimates are not informative for predicting the long-term effect of horizontal gene transfer in bacteria

Horizontal gene transfer (HGT) is an important mechanism by which bacteria recombine and acquire novel genes and functions. Risk scenarios where novel plant transgenes transfer horizontally into bacteria have been addressed in numerical theoretical assessments and experimental studies. A key outcome of these studies has been that the frequencies of such inter-domain transfer are very low, if occurring at all, suggesting that such transfers would not occur at a level that is biologically significant. The relationship between transfer frequencies and the subsequent selection or genetic drift of transgene carrying bacteria often remains unresolved in these studies and assessments. Here we present a stochastic model to better understand the initial establishment and population dynamics of rare bacterial transformants carrying horizontally acquired (trans)genes. The following key parameters are considered: initial transformant numbers, strength of selection, bacterial population size and bacterial generations (time). We find that the initial number of transformants is important for the subsequent persistence of transformants only in the range of 1 to approximately 50 independent HGT events. Our simulations show that transformant populations under a wide range of HGT rates and selection coefficients undergo stochastic developments where they persist at low frequencies for up to several years (at frequencies that are below detection using available field sampling methodology), after which they eventually may go to fixation. Stochastic variability may thus play a crucial but disregarded role in the design of field monitoring strategies e.g. in biosafety assessments. We also estimate the time required for transformants to reach 0.0002% prevalence in a bacterial population, a threshold that allows experimental detection of transgene carrying bacteria through sampling of the larger bacterial populations.

Eubacterial communities in different soil macroaggregate environments and cropping systems

Different positions within soil macroaggregates, and macroaggregates of different sizes, have different chemical and physical properties which could affect microbial growth and interactions among taxa. The hypothesis that these soil aggregate fractions contain different eubacterial communities was tested using terminal restriction fragment length polymorphism (T-RFLP) of the 16S ribosomal gene. Communities were characterized from two field experiments, located at the Kellogg Biological Station (KBS), MI, USA and the Ohio Agricultural Research and Development Center (OARDC), Wooster, OH, USA. Three soil management regimes at each site were sampled and management was found to significantly affect T-RFLP profiles. The soil aggregate erosion (SAE) method was used to isolate aggregate regions (external and internal regions). Differences between eubacterial T-RFLP profiles of aggregate exteriors and interiors were marginally significant at KBS (accounting for 12.5% of total profile variance), and not significant at OARDC. There were no significant differences among macroaggregate size classes at either site. These results are in general agreement with previous studies using molecular methods to examine microbial communities among different soil macroaggregate size fractions, although further study of communities within different aggregate regions is warranted. Analysis of individual macroaggregates revealed large inter-aggregate variability in community structure. Hence the tertiary components of soil structure, e.g. arrangement of aggregates in relation to shoot residue, roots, macropores, etc., may be more important than aggregate size or intra-aggregate regions in the determination of the types of microbial communities present in aggregates. Direct microscopic counts were also used to examine the bacterial population size in aggregate regions at KBS. The proportion of bacterial cells with biovolumes >0.18 μm 3 was higher in aggregate interiors than in exteriors, indicating potentially higher activity in that environment. This proportion was significantly related to percent C of the samples, while total bacterial cell counts were not.

Assessing interactions of multiple agrichemicals by using bacterial assemblages in a wetland mesocosm system.

Agrichemicals may enter wetlands located adjacent to or downstream from agricultural fields. We investigated the individual and interactive effects of three agrichemicals [atrazine, chlorpyrifos, and monosodium acid methanearsonate (MSMA)] and methyl mercury on abundance and heterotrophic potential of wetland heterotrophic bacteria assemblages. We used a factorial experimental design, in which chemicals were introduced in all possible combinations to 66 500-liter mesocosms at the Biological Field Station of the University of Mississippi. Methyl mercury was added to bring the total mercury (HG) concentration to 0.4 mg/Kg wet weight at the sediment surface. Atrazine, chlorpyrifos, and MSMA were added at concentrations of 192, 51, and 219 microg/L, respectively. Over 32 days of exposure, microbial heterotrophic activity was sensitive to only the interactive effect of HG*ATR*CPF in the sediments and only CPF in the water. Total bacterial numbers did not exhibit any significant treatment effects. Therefore, the effects of agrichemicals were reflected on cell-specific bacterial heterotrophic activity rather than bacterial population size.

Bacterial Colonization of the Phyllosphere of Mediterranean Perennial Species as Influenced by Leaf Structural and Chemical Features

In this study, we assessed various leaf structural and chemical features as possible predictors of the size of the phyllosphere bacterial population in the Mediterranean environment. We examined eight perennial species, naturally occurring and coexisting in the same area, in Halkidiki (northern Greece). They are Arbutus unedo, Quercus coccifera, Pistacia lentiscus, and Myrtus communis (evergreen sclerophyllous species), Lavandula stoechas and Cistus incanus (drought semi-deciduous species), and Calamintha nepeta and Melissa officinalis (non-woody perennial species). M. communis, L. stoechas, C. nepeta, and M. officinalis produce essential oil in substantial quantities. We sampled summer leaves from these species and (1) estimated the size of the bacterial population of their phyllosphere, (2) estimated the concentration of different leaf constituents, and (3) studied leaf morphological and anatomical features and expressed them in a quantitative way. The aromatic plants are on average more highly colonized than the other species, whereas the non-woody perennials are more highly colonized than the woody species. The population size of epiphytic bacteria is positively correlated with glandular and non-glandular trichome densities, and with water and phosphorus contents; it is negatively correlated with total phenolics content and the thickness of the leaf, of the mesophyll, and of the abaxial epidermis. No correlation was found with the density of stomata, the nitrogen, and the soluble sugar contents. By regression tree analysis, we found that the leaf-microbe system can be effectively described by three leaf attributes with leaf water content being the primary explanatory attribute. Leaves with water content >73% are the most highly colonized. For leaves with water content <73%, the phosphorus content, with a critical value of 1.34 mg g(-1) d.w., is the next explanatory leaf attribute, followed by the thickness of the adaxial epidermis. Leaves higher in phosphorus (>1.34 mg g(-1) d.w.) are more colonized, and leaves with the adaxial epidermis thicker than 20.77 microm are the least colonized. Although these critical attributes and values hold true only within the Mediterranean ecosystem studied and the range of observations taken, they are important because they provide a hypothesis to be tested in other Mediterranean ecosystems and other biomes. Such comparative studies may give insight as to the general properties governing the leaf-microbe system.

A Stochastic Formulation of the Gompertzian Growth Model forin vitro Bactericidal Kinetics: Parameter Estimation and Extinction Probability

Time-kill curves have frequently been employed to study the antimicrobial effects of antibiotics. The relevance of pharmacodynamic modeling to these investigations has been emphasized in many studies of bactericidal kinetics. Stochastic models are needed that take into account the randomness of the mechanisms of both bacterial growth and bacteria-drug interactions. However, most of the models currently used to describe antibiotic activity against microorganisms are deterministic. In this paper we examine a stochastic differential equation representing a stochastic version of a pharmacodynamic model of bacterial growth undergoing random fluctuations, and derive its solution, mean value and covariance structure. An explicit likelihood function is obtained both when the process is observed continuously over a period of time and when data is sampled at time points, as is the custom in these experimental conditions. Some asymptotic properties of the maximum likelihood estimators for the model parameters are discussed. The model is applied to analyze in vitro time-kill data and to estimate model parameters; the probability of the bacterial population size dropping below some critical threshold is also evaluated. Finally, the relationship between bacterial extinction probability and the pharmacodynamic parameters estimated is discussed.

Genetic architecture of Arabidopsis thaliana response to infection by Pseudomonas syringae

Plant pathogens can severely reduce host yield and fitness. Thus, investigating the genetic basis of plant response to pathogens is important to further understand plant-pathogen coevolution and to improve crop production. The interaction between Arabidopsis thaliana and Pseudomonas syringae is an important model for studying the genetic basis of plant-pathogen interactions. Studies in this model have led to the discovery of many genes that differentiate a resistant from a susceptible plant. However, little is known about the genetic basis of quantitative variation in response to P. syringae. In this study, we investigate the genetic basis of three aspects of A. thaliana's response to P. syringae: symptom severity, bacterial population size and fruit production using a quantitative trait loci (QTL) analysis. We found two QTL for symptom severity and two for fruit production (possible candidate genes for observed QTL are discussed). We also found significant two-locus epistatic effect on symptom severity and fruit production. Although bacterial population size and symptom severity were strongly phenotypically correlated, we did not detect any QTL for bacterial population size. Despite the detected genetic variation observed for susceptibility, we found only a weak overall relationship between susceptibility traits and fitness, suggesting that these traits may not respond to selection.

Quantification of Ammonia-Oxidizing Bacteria and Factors Controlling Nitrification in Salt Marsh Sediments

To elucidate the geomicrobiological factors controlling nitrification in salt marsh sediments, a comprehensive approach involving sediment geochemistry, process rate measurements, and quantification of the genetic potential for nitrification was applied to three contrasting salt marsh habitats: areas colonized by the tall (TS) or short (SS) form of Spartina alterniflora and unvegetated creek banks (CBs). Nitrification and denitrification potential rates were strongly correlated with one another and with macrofaunal burrow abundance, indicating that coupled nitrification-denitrification was enhanced by macrofaunal burrowing activity. Ammonia monooxygenase (amoA) gene copy numbers were used to estimate the ammonia-oxidizing bacterial population size (5.6 x 10(4) to 1.3 x 10(6) g of wet sediment(-1)), which correlated with nitrification potentials and was 1 order of magnitude higher for TS and CB than for SS. TS and CB sediments also had higher Fe(III) content, higher Fe(III)-to-total reduced sulfur ratios, higher Fe(III) reduction rates, and lower dissolved sulfides than SS sediments. Iron(III) content and reduction rates were positively correlated with nitrification and denitrification potential and amoA gene copy number. Laboratory slurry incubations supported field data, confirming that increased amounts of Fe(III) relieved sulfide inhibition of nitrification. We propose that macrofaunal burrowing and high concentrations of Fe(III) stimulate nitrifying bacterial populations, and thus may increase nitrogen removal through coupled nitrification-denitrification in salt marsh sediments.

Experimental adaptation of Salmonella typhimurium to mice.

Experimental evolution is a powerful approach to study the dynamics and mechanisms of bacterial niche specialization. By serial passage in mice, we evolved 18 independent lineages of Salmonella typhimurium LT2 and examined the rate and extent of adaptation to a mainly reticuloendothelial host environment. Bacterial mutation rates and population sizes were varied by using wild-type and DNA repair-defective mutator (mutS) strains with normal and high mutation rates, respectively, and by varying the number of bacteria intraperitoneally injected into mice. After <200 generations of adaptation all lineages showed an increased fitness as measured by a faster growth rate in mice (selection coefficients 0.11-0.58). Using a generally applicable mathematical model we calculated the adaptive mutation rate for the wild-type bacterium to be >10(-6)/cell/generation, suggesting that the majority of adaptive mutations are not simple point mutations. For the mutator lineages, adaptation to mice was associated with a loss of fitness in secondary environments as seen by a reduced metabolic capability. During adaptation there was no indication that a high mutation rate was counterselected. These data show that S. typhimurium can rapidly and extensively increase its fitness in mice but this niche specialization is, at least in mutators, associated with a cost.

Distribution of Xylella fastidiosa in Sycamore Associated with Low Temperature and Host Resistance.

Experiments were conducted in the field and laboratory to determine effects of low temperatures on Xylella fastidiosa populations in American sycamore. Roots and shoots from naturally infected trees at two locations were collected monthly. Sap extracted from the samples was tested by enzyme-linked immunosorbent assay for presence of X. fastidosa and was diluted and plated on periwinkle wilt medium to determine populations of viable bacteria. Cumulative rainfall and hours below temperature thresholds (-5 to 10°C) were recorded at each site. Bacterial populations in shoots were negatively correlated with cumulative hours below -5°C air temperature (r= -0.96). In roots, bacterial populations were only weakly correlated with cumulative hours below soil temperature thresholds (-0.61 < r <-0.25). Bacterial populations were not correlated with monthly rainfall. In the laboratory, resistant and susceptible sycamore trees were inoculated with X. fastidiosa and held in the dark at 5°C or 22°C. After 12 weeks, inoculated stem sections were collected and sap was extracted and tested as described previously. Stems that tested positive for X. fastidiosa were divided into additional samples and tested as described above. Results of the laboratory study indicated no significant effects of low-temperature treatment (5°C) or host resistance on viable bacteria. Bacterial detection frequency and population size were greatest near the inoculation point and the primary direction of early bacterial spread was acropetal.

Advertising as Public Diplomacy: Attitude Change among International Audiences

1.<h3>ABSTRACT</h3> <h3>Background</h3> Drug-resistant tuberculosis is a high priority threat to global public health. There are still critical gaps in understanding how novel drug-resistant <i>M. tuberculosis</i> strains emerge and, once emergent, what drives the differential propagation of certain epidemiologically-successful strains over others. This study sought to describe the joint evolutionary and epidemiological histories of a novel multidrug-resistant <i>M. tuberculosis</i> strain recently identified in the capital city of the Republic of Moldova (MDR Ural/4.2). <h3>Methods</h3> Using whole genome sequence data and Bayesian phylogenomic methods, we reconstruct the stepwise acquisition of drug-resistance mutations in the MDR Ural/4.2 strain, estimate its historical bacterial population size over time, and infer the migration history of this strain between Eastern European countries. <h3>Results</h3> We infer that MDR Ural/4.2 likely evolved (via acquisition of <i>rpoB</i> S450L, which confers resistance to rifampin) in the early 1990s, during a period of social turmoil following Moldovan independence from the Soviet Union. This strain subsequently underwent substantial population size expansion in the early 2000s, at a time when national guidelines encouraged in hospital treatment of TB patients. We infer exportation of this strain and its INH-resistant ancestral precursor from Moldova to neighboring countries starting as early as 1985. <h3>Conclusions</h3> Our findings underscore how public health practice and social determinants of health shape the conditions under which <i>M. tuberculosis</i> evolves, and demonstrates how historical changes in these conditions shape present-day challenges in TB control. These findings underscore the need for regional coordination in TB control across Eastern Europe.

Influence of temperature on plant–rhizobacteria interactions related to biocontrol potential for suppression of fusarium wilt of chickpea

Seed and soil treatment with Pseudomonas fluorescens RGAF 19, P. fluorescens RG 26, Bacillus megaterium RGAF 51 and Paenibacillus macerans RGAF 101 can suppress fusarium wilt of chickpea (Cicer arietinum), but the extent of disease suppression by these rhizobacteria is modulated by soil temperature. In this work, the effect of temperature on plant–rhizobacteria interactions was assessed in relation to biocontrol potential for suppression of fusarium wilt of chickpea. Seed and soil treatment with those rhizobacteria delayed seedling emergence compared with nontreated controls, and either increased or had no deleterious effect on chickpea growth. Pseudomonas fluorescens isolates significantly increased chickpea shoot dry weight at 20°C and root dry weight at 25 and 30°C. All bacterial isolates colonized the chickpea rhizosphere and internal stem tissues at 20, 25 and 30°C, and there was a positive linear trend between bacterial population size in the rhizosphere and temperature increase. The maximum inhibition of mycelial growth and conidial germination of Fusarium oxysporum f. sp. ciceris race 5 in vitro occurred at a temperature range optimal for bacterial growth and production of inhibitory metabolites. These results demonstrate the need to understand the effects of environmental factors on the biological activities of introduced rhizobacteria of significant importance for plant disease suppression.

Influence of Antimicrobial Agents on the Spoilage of a Meat-Based Entomophage Diet

The microbial decomposition of a meat-based entomophage diet presented in Parafilm packets was investigated. Considerable bacteria but not fungi were associated with components used to prepare the diet (i.e., hens’ eggs, liver, and ground beef). At the initial sampling time, there were no differences among diet treatments in the size of bacterial or fungal populations. Bacterial populations in diets not containing antibacterial agents rapidly increased and reached an asymptote by 24 h (≈1010 colony-forming units per gram). Bacterial populations also increased in diets containing antibacterial agents, but they were significantly smaller than in diets not containing antibacterial agents. The most prevalent bacteria isolated were Carnobacterium piscicola, Carnobacterium divergens, Lactobacillus curvatus, Lactobacillus sakei, Leuconostoc mesenteroides, and Enterococcus spp., regardless of the antibacterial treatment used. The proliferation of fungi was delayed relative to bacteria, but significant differences were observed among the diet treatments. Fungi were most inhibited by sorbic acid and propionic acid in the absence of antibacterial agents. The most common fungi isolated were the yeasts Candida zeylanoides, Torulaspora globosa, and Yarrowia lipolytica. The pH of diets not containing antibacterial agents decreased rapidly and was highly correlated with increases in bacteria but not fungi. The results of this study demonstrate that antimicrobial agents significantly inhibit spoilage microorganisms in a meat-based diet and that alternative management strategies to delay the decomposition of such diets presented in Parafilm packets should target lactic acid spoilage bacteria, particularly Carnobacterium and Lactobacillus species.

Influence of Antimicrobial Agents on the Spoilage of a Meat-Based Entomophage Diet

The microbial decomposition of a meat-based entomophage diet presented in Parafilm packets was investigated. Considerable bacteria but not fungi were associated with components used to prepare the diet (i.e., hens' eggs, liver, and ground beef). At the initial sampling time, there were no differences among diet treatments in the size of bacterial or fungal populations. Bacterial populations in diets not containing antibacterial agents rapidly increased and reached an asymptote by 24 h (approximately 10(10) colony-forming units per gram). Bacterial populations also increased in diets containing antibacterial agents, but they were significantly smaller than in diets not containing antibacterial agents. The most prevalent bacteria isolated were Carnobacterium piscicola, Carnobacterium divergens, Lactobacillus curvatus, Lactobacillus sakei, Leuconostoc mesenteroides, and Enterococcus spp., regardless of the antibacterial treatment used. The proliferation of fungi was delayed relative to bacteria, but significant differences were observed among the diet treatments. Fungi were most inhibited by sorbic acid and propionic acid in the absence of antibacterial agents. The most common fungi isolated were the yeasts Candida zeylanoides, Torulaspora globosa, and Yarrowia lipolytica. The pH of diets not containing antibacterial agents decreased rapidly and was highly correlated with increases in bacteria but not fungi. The results of this study demonstrate that antimicrobial agents significantly inhibit spoilage microorganisms in a meat-based diet and that alternative management strategies to delay the decomposition of such diets presented in Parafilm packets should target lactic acid spoilage bacteria, particularly Carnobacterium and Lactobacillus species.

Pseudomonas syringae pv. tomato cells encounter inhibitory levels of water stress during the hypersensitive response of Arabidopsis thaliana.

During plant defense against bacterial pathogens, the hypersensitive response (HR) functions to restrict pathogen growth and spread. The mechanisms driving this growth restriction are poorly understood. We used a water stress-responsive transcriptional fusion to quantify the water potential sensed by individual Pseudomonas syringae pv. tomato DC3000 cells during infection of Arabidopsis thaliana leaves. A nonpathogenic DC3000 hrcC mutant defective in type III secretion, as well as the saprophyte Pseudomonas fluorescens A506, sensed water potentials of -0.3 to -0.4 MPa at 48 h postinfiltration (hpi). During pathogenesis, DC3000 sensed lower water potentials (-0.4 to -0.9 MPa), demonstrating that it can modify the intercellular environment, and these water potentials were associated with optimal DC3000 growth in culture. During the HR, DC3000 cells sensed water potentials (-1.6 to -2.2 MPa) that were low enough to prevent cell division in the majority of cells in culture. This water potential decrease occurred within only 4 hpi and was influenced by avirulence gene expression, with avrRpm1 expression associated with lower water potentials than avrRpt2 or avrB expression at 48 hpi. The population sizes of the DC3000 variants tested were significantly correlated with the apoplastic water potential at 48 hpi, with a decrease of -0.9 MPa associated with a 10-fold decrease in cells per gram of leaf. These results suggest that the apoplastic water potential is a determinant of endophytic bacterial population size, and water stress, resulting from high osmolarity or tissue desiccation, is at least one factor restricting bacterial growth during the HR.

INVESTIGATION OF THE RELIABILITY OF EASY-TO-USE METHODS TO PREDICT ERWINIA AMYLOVORA INFECTION RISK IN APPLE ORCHARDS

As a complement to a national fire blight monitoring network, such as the MARYBLYT system in Hungary, an accurate local prediction method would improve control in a large orchard or a micro-region. Data relating to temperature and wetting events in 1999-2001 were analysed by means of the Californian Mean temperature line, Smith's degree hour-based model (Cougarblight), Billing's integrated system (BIS95) and the MARYBLYT computer model. In two years, the start of the massive infection risk was predicted by three of four models on the same day: 28 April in 1999 and 25 April in 2000. However, there was a delay of one day with the Cougarblight model as compared with the BIS95 and MARYBLYT systems in 2001. The forecast of an infection risk via the Mean Temperature Line was consequently one day later than that with the BIS95 and MARYBLYT systems. The size of the epiphytic bacterial population in the flowers was variable from year to year, with no convincing difference between cultivars with high (e.g. 'Sampion') or low (e.g. 'Freedom') susceptibility. The incidence of fire blight was sporadic in 1999 and 2001, but epidemic in 2000. The proportion of trees of 6 apple cultivars with fire blight symptoms well reflected the differences in weather-related risks in the two years investigated. The findings indicate that both Cougarblight and BIS95 deserve to be tested in commercial orchards in Hungary.

Genetic variation for disease resistance and tolerance among Arabidopsis thaliana accessions.

Pathogens can be an important selective agent in plant evolution because they can severely reduce plant fitness and growth. However, the role of pathogen selection on plant evolution depends on the extent of genetic variation for resistance traits and their covariance with host fitness. Although it is usually assumed that resistance traits will covary with plant fitness, this assumption has not been tested rigorously in plant-pathogen interactions. Many plant species are tolerant to herbivores, decoupling the relationship between resistance and fitness. Tolerance to pathogens can reduce selection for resistance and alter the effect of pathogens on plant evolution. In this study, we measured three components of Arabidopsis thaliana resistance (pathogen growth, disease symptoms, and host fitness) to the bacteria Pseudomonas syringae and investigated their covariation to determine the relative importance of resistance and tolerance. We observed extensive quantitative variation in the severity of disease symptoms, the bacterial population size, and the effect of infection on host fitness among 19 accessions of A. thaliana infected with P. syringae. The severity of disease symptoms was strongly and positively correlated with bacterial population size. Although the average fitness of infected plants was smaller than noninfected plants, we found no correlation between the bacterial growth or symptoms expressed by different accessions of A. thaliana and their relative fitness after infection. These results indicate that the accessions studied vary in tolerance to P. syringae, reducing the strength of selection on resistance traits, and that symptoms and bacterial growth are not good predictors of host fitness.

The Effect of NaCl on Survival of Shigella flexneri in Broth as Affected by Temperature and pH

Shigella, a major foodborne pathogen, survives well in salt-containing environments. However, systematic data are scarce. We studied the behavior of Shigella flexneri 5348 in brain heart infusion broth (pH 4 to 6) containing 0.5 to 8% NaCl. Stationary-phase cells were inoculated into sterile media at initial concentrations of 6 to 7 log10 CFU/ml and incubated at 12 to 37°C. Bacterial population sizes were determined periodically by plate counts. Survivor curves were derived from plate count data by using a two-phase linear model to determine lag times and slopes of the curves, from which decimal reduction times (D-values) and times to a 4-log10 inactivation (T4D) were calculated. In media of pH 6, the bacteria grew in the presence of ≤6% NaCl at 19 and 37°C and in the presence of ≤7% NaCl at 28°C. In media of pH 5, growth was observed in the presence of ≤2, ≤4, ≤4, and 0.5% NaCl at 37, 28, 19, and 12°C, respectively. Growth did not occur and bacterial populations gradually declined in media of pH 4. While NaCl had a major effect on growth, bacterial survival was affected to a lesser extent. Lag times decreased with increasing NaCl levels; however, the effect on D-values and T4D values was less pronounced. The average T4D values for media of pH 4 containing 0.5 to 6% NaCl were 4, 13, 23, and 61 days at 37, 28, 19, and 12°C, respectively. These results show that S. flexneri is salt tolerant and suggest that salty foods may serve as vehicles for infection with this bacterium.