scholarly journals Growth and extended survival ofEscherichia coliO157:H7 in soil organic matter

2017 ◽  
Author(s):  
Gitanjali NandaKafle ◽  
Amy A. Christie ◽  
Sébastien Vilain ◽  
Volker S. Brözel
Keyword(s):  
Organic Matter ◽  
Stationary Phase ◽  
Open Systems ◽  
Physiological State ◽  
Principal Component ◽  
Human Infection ◽  
Food Plants ◽  
Bacterial Populations ◽  
E Coli ◽  
2D Gel

AbstractEnterohaemorrhagicEscherichia colisuch as serotype O157:H7 are a leading cause of food-associated outbreaks. While the primary reservoir is associated with cattle, plant foods have been associated as sources of human infection.E. coliis able to grow in the tissue of food plants such as spinach. While fecal contamination is the primary suspect, soil has been underestimated as a potential reservoir. Persistence of bacterial populations in open systems is the product of growth, death, predation, and competition. Here we report thatE. coliO157:H7 can grow using the soluble compounds in soil, and characterize the effect of soil growth in the stationary phase proteome.E. coli933D (stxII-) was cultured in Soil Extracted Soluble Organic Matter (SESOM) and the culturable count determined for 24 d. The proteomes of exponential and stationary phase populations were characterized by 2D gel electrophoresis and protein spots were identified by MALDI-TOF mass spectrometry. While LB controls displayed a death phase, SESOM grown population remained culturable for 24 d, indicating an altered physiological state with superior longevity. This was not due to decreased cell density on entry to stationary phase as 24h SESOM populations concentrated 10-fold retained their longevity. Principal component analysis showed that stationary phase proteomes from SESOM and LB were different. Differences included proteins involved in stress response, motility, membrane and wall composition, nutrient uptake, translation and protein turnover, and anabolic and catabolic pathways, indicating an altered physiological state of soil-grown cells entering stationary phase. The results suggest thatE. colimay be a soil commensal that in absence of predation and competition maintains stable populations in soil.

Effects of synthetic organic chemicals on bacterial death and lysis

1998 ◽  
Vol 37 (4-5) ◽  
pp. 385-393 ◽  
Author(s):  
G. G. Perez-Padilla ◽  
C. P. L. Grady
Keyword(s):  
Stationary Phase ◽  
Organic Compounds ◽  
Tritiated Thymidine ◽  
Physiological State ◽  
Radioactive Tracer ◽  
Major Effect ◽  
Specific Rate ◽  
E Coli ◽  
Lysis Rate ◽  
Synthetic Organic Compounds

A technique derived from the tritiated thymidine method was used to characterize the effects of synthetic organic compounds (SOCs) on the specific rate of bacterial death as functions of chemical concentration, the physiological state of the cells, and exposure time to the toxicant. The rate of bacterial death was estimated by following over time the release of radioactive tracer from the DNA of thymidine-requiring (thy−) mutant E. coli cells (ATCC 23820). Results indicate that the lysis rate of unexposed microbial cultures was the same whether or not the cells were growing or had reached the stationary phase. Lysis rates were calculated from exposures to single SOCs: acrylonitrile, ethylene glycol, isophorone, phenol, 2-chloro- and 4-chlorophenol. The concentrations tested were 250, 1000 and 5000 mg COD/l. The major effect observed with stationary-phase cultures was a stimulation of the rate of lysis, whereas a reduction in the rate of lysis was the primary effect observed with growing cells. A physiological interpretation for these opposite effects is provided. The physiological state of the microorganisms influenced the magnitude, intensity and type of effects caused by the presence of synthetic organic compounds.

scholarly journals Transcriptome signature of E. coli under a prolonged starvation environment

2020 ◽  
Author(s):  
Sotaro Takano ◽  
Hiromi Takahashi ◽  
Yoshie Yama ◽  
Ryo Miyazaki ◽  
Saburo Tsuru
Keyword(s):  
Stationary Phase ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Physiological State ◽  
Exponential Growth Phase ◽  
Transcriptional Responses ◽  
E Coli ◽  
Viable Cells ◽  
Genome Wide

ABSTRACTBackground“Non-growing” is a dominant life form of microorganisms in nature, where available nutrients and resources are extremely limited. However, the knowledge of the manner in which microorganisms resist nutrient deficiency is still rudimentary compared to those of the growing cells. In laboratory culture, Escherichia coli can survive for several years under starvation, denoted as long-term stationary phase (LSP), where a small fraction of the cells survive by recycling resources released from the starved nonviable cells and constitute a model system for understanding survival mechanisms under long-term starvation. Although the physiology by which viable cells in LSP adapt to long-term starvation is of great interest, their genome-wide response has not yet been fully understood.ResultsTo understand the physiological state of viable cells in the LSP environment, we analyzed the transcriptional profiles of cells exposed to the supernatant of LSP culture. We found that high expression of transporter genes and low expression of biosynthesis genes are the primary responses of the cells in the LSP supernatant compared to growing cells, which display similar responses to cells entering the stationary phase from the exponential growth phase. We also revealed some specific transcriptional responses in the LSP supernatant, such as higher expression of stress-response genes and lower expression of translation-related genes, compared to other non-growing conditions. This suggests that cells in LSP are highly efficient in terms of cellular survival and maintenance functions under starvation conditions. We also found population-density-dependent gene expression profiles in LSP, which are also informative to understand the survival mechanism of bacterial population.ConclusionOur current comprehensive analysis of the transcriptome of E. coli cells provides an overview of the genome-wide response to the long-term starvation environment. We detected both common and distinctive responses in the primary transcriptional changes between the short- and long-term stationary phase cultures, which could provide clues to understand the possible molecular mechanisms underlying survivability in the starved environment.

Antecedent Oxygen Growth Conditions and Recovery of Heat-Stressed Escherichia coli1

1991 ◽  
Vol 54 (2) ◽  
pp. 90-93 ◽  
Author(s):  
CAROLINE E. O'NEILL ◽  
GARY K. BISSONNETTE
Keyword(s):  
Escherichia Coli ◽  
Heat Stress ◽  
Thermal Stress ◽  
Stationary Phase ◽  
Physiological State ◽  
Growth Conditions ◽  
Exponential Phase ◽  
Physiological Age ◽  
E Coli

Four strains of Escherichia coli were examined for response to heat stress (60°C) as a function of physiological age and antecedent oxygen growth conditions. Exponential phase cells were more susceptible to heat than cells grown to the stationary phase. Anaerobically grown, exponential phase cells were more susceptible to thermal stress than were cells grown to a similar physiological state but under aerobic conditions. In the case of stationary phase cells, differences in response to heat stress as related to prior oxygen growth conditions were equivocal. Repair characteristics of thermally injured cells were also examined. Cells grown anaerobically prior to heat stress required 1.5 h longer than their aerobically grown counterparts to complete repair. These findings suggest that antecedent oxygen growth conditions influence the response of E. coli to thermal stress and perhaps, more generally, that persistence of environmentally stressed enteric microorganisms must be considered in relation to prior oxygen growth conditions in vivo.

scholarly journals Fluorescent reporter plasmids for single-cell and bulk-level composition assays in E. faecalis

2019 ◽  
Author(s):  
Kelsey M. Hallinen ◽  
Keanu A. Guardiola-Flores ◽  
Kevin B. Wood
Keyword(s):  
Stationary Phase ◽  
Single Cell ◽  
Opportunistic Pathogen ◽  
Spectral Unmixing ◽  
Model Organisms ◽  
Fluorescent Reporter ◽  
Bacterial Populations ◽  
Fluorescent Reporters ◽  
E Coli ◽  
Scope Of Application

ABSTRACTFluorescent reporters are an important tool for monitoring dynamics of bacterial populations at the single cell and community level. While there are a large range of reporter constructs available–particularly for common model organisms like E. coli–fewer options exist for other species, including E. faecalis, a gram-positive opportunistic pathogen. To expand the potential toolkit available for E. faecalis, we modified a previously developed reporter plasmid (pBSU101) to express one of nine different fluorescent reporters and confirmed that all constructs exhibited detectable fluorescence in single E. faecalis cells and mixed biofilm communities. To identify promising constructs for bulk-level experiments, we then measured the fluorescence spectra from E. faecalis populations in microwell plate (liquid) cultures during different growth phases. Cultures showed density- and reporter-specific variations in fluorescent signal, though spectral signatures of all reporters become clear in late-exponential and stationary-phase populations. Based on these results, we identified six pairs of reporters that can be combined with simple spectral unmixing to accurately estimate population composition in 2-strain mixtures at or near stationary phase. This approach offers a simple and scalable method for selection and competition experiments in simple two-species populations. Finally, we modified the construct to express codon-optimized variants of blue (BFP) and red (RFP) reporters and show that they lead to increased fluorescence in exponentially growing cells. As a whole, the results inform the scope of application of different reporters and identify both single reporters and reporter pairs that are promising for fluorescence-based assays at bulk and single-cell levels in E. faecalis.

In vitro antibacterial and antibiotic-potentiation activities of five edible plant extracts and mode of action against several MDR Gram-negative phenotypes

2020 ◽  
Vol 4 (1) ◽  
pp. 1-14
Author(s):  
Carine M.N. Ngaffo ◽  
Simplice B. Tankeo ◽  
Michel-Gael F. Guefack ◽  
Brice E. N. Wamba ◽  
Paul Nayim ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Bioactive Compounds ◽  
Plant Extracts ◽  
Theobroma Cacao ◽  
Efflux Pumps ◽  
Phoenix Dactylifera ◽  
Food Plants ◽  
Proton Pumps ◽  
Gram Negative ◽  
E Coli

Abstract Background: Bacterial infections involving the multidrug resistant (MDR) strains are among the top leading causes of death throughout the world. Healthcare system across the globe has been suffering from an extra-ordinary burden in terms of looking for the new and more potent antimicrobial compounds. The aim of the present study was to determine the antibacterial activity of some Cameroonian edible plants (Garcinia lucida bark, Phoenix dactylifera pericarps, Theobroma cacao pod, Solanum macrocarpon leaves and Termitomyces titanicus whole plant) and their antibiotics-potentiation effects against some MDR Gram-negative bacteria phenotypes expressing efflux pumps (Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas aeruginosa and Providencia stuartii strains). Methods: The antibacterial activities of plant extract alone and in combination with usual antibiotics were carried out using the micro-dilution method. The effects of the most active plant extract (Garcinia lucida bark) on H+-ATPase-mediated proton pumps and on bacterial growth kinetic were performed using experimental protocols, while qualitative reference methods were used to highligh the major groups of secondary metabolites present in the extracts. Results: Qualitative phytochemical screening of plant extracts indicated that all analysed secondary metabolites were present in Theobroma cacao and Termitomyces titanicus while one (saponins) of them was absent in Garcinia lucida and Solanum macrocarpon. Only three of them (polyphenols, flavonoids and saponins) were detected in Phoenix dactylifera. Antibacterial essays showed that G. lucida was the most active plant as it inhibited the growth of all studied bacteria with strong activity (MIC<100 µg/mL) against E. coli ATCC8739, significant activity (100≤MIC≤512 µg/mL) against 80% of bacteria and moderate activity (512<MIC≤2048 µg/mL) against E. coli AG100A and E. aerogenes (EA289 and CM64). It was followed by T. cacao and S. macrocarpon extracts which exhibited an antibacterial potential against 95% and 80% of bacterial strains, respectively. These three extracts exhibited a bactericidal effect on a few bacteria. Extracts from T. titanicus and P. dactylifera were less active as they moderately (512<MIC≤2048 µg/mL) inhibited the growth of 35% and 10% of bacteria. All extracts selectively potentiated the activities of all antibiotics with improvement activity factors (IAF) ranging from 2 to 256. G. lucida, T. cacao and S. macrocarpon potentiated the activities of 100%, 89% and 67% of antibiotics respectively against more than 70%, suggesting that they contain bioactive compounds which could be considered as efflux pumps inhibitors. Whereas T. titanicus and P. dactylifera improved the activities of almost 40% and 20% of antibiotics, respectively. This increase of activities also characterizes synergistic effects between antibiotics and these bioactive compounds. G. lucida extract at all tested concentrations, strongly inhibited the growth of bacterial strain E. coli ATCC8739 and exhibited an inhibitory effect on this bacterial H+-ATPase-mediated proton pumps increasing the pH of the medium. Conclusion: The overall results indicated that food plants among which G. lucida, T. cacao and S. macrocarpon could have a benefit interest in combatting resistant types of bacteria. Keywords: Food plants; infectious diseases; MDR bacteria; efflux pumps; antibiotics; secondary metabolites.

scholarly journals Impact of aquaculture on distribution of dissolved organic matter in coastal Jeju Island, Korea, based on absorption and fluorescence spectroscopy

2021 ◽  
Author(s):  
Jeonghyun Kim ◽  
Yeseul Kim ◽  
Sung Eun Park ◽  
Tae-Hoon Kim ◽  
Bong-Guk Kim ◽  
...  
Keyword(s):  
Organic Matter ◽  
Organic Carbon ◽  
Fluorescence Spectroscopy ◽  
Dissolved Organic Matter ◽  
Coastal Water ◽  
Coastal Region ◽  
Principal Component ◽  
Distribution Patterns ◽  
Organic Pollution ◽  
Jeju Island

AbstractIn Jeju Island, multiple land-based aquafarms were fully operational along most coastal region. However, the effect of effluent on distribution and behaviours of dissolved organic matter (DOM) in the coastal water are still unknown. To decipher characteristics of organic pollution, we compared physicochemical parameters with spectral optical properties near the coastal aquafarms in Jeju Island. Absorption spectra were measured to calculate the absorption coefficient, spectral slope coefficient, and specific UV absorbance. Fluorescent DOM was analysed using fluorescence spectroscopy coupled with parallel factor analysis. Dissolved organic carbon (DOC) and total dissolved nitrogen (TDN) were measured using high-temperature catalytic oxidation. The DOC concentration near the discharge outlet was twice higher than that in natural groundwater, and the TDN concentration exponentially increased close to the outlet. These distribution patterns indicate that aquafarms are a significant source of DOM. Herein, principal component analysis was applied to categorise the DOM origins. There were two distinct groups, namely, aquaculture activity for TDN with humic-like and high molecular weights DOM (PC1: 48.1%) and natural biological activity in the coastal water for DOC enrichment and protein-like DOM (PC2: 18.8%). We conclude that the aquafarms significantly discharge organic nitrogen pollutants and provoke in situ production of organic carbon. Furthermore, these findings indicate the potential of optical techniques for the efficient monitoring of anthropogenic organic pollutants from aquafarms worldwide.

scholarly journals Effect of Environmental Factors and Influence of Rumen and Hindgut Biogeography on Bacterial Communities in Steers

2010 ◽  
Vol 77 (1) ◽  
pp. 258-268 ◽  
Author(s):  
Gustavo A. Romero-Pérez ◽  
Kim H. Ominski ◽  
Tim A. McAllister ◽  
Denis O. Krause
Keyword(s):  
Ambient Temperature ◽  
Bacterial Communities ◽  
Volatile Fatty Acids ◽  
Principal Component ◽  
Population Diversity ◽  
Access Time ◽  
Sample Type ◽  
Bacterial Populations ◽  
Fecal Samples ◽  
Alfalfa Silage

ABSTRACTFeces from cattle production are considered important sources of bacterial contamination of food and the environment. Little is known about the combined effects of arctic temperatures and fodder tannins on rumen and hindgut bacterial populations. Individual rumen liquor and rectal fecal samples from donor steers fed either alfalfa silage or sainfoin (Onobrychis viciifoliaScop.) silage and waterad libitumwere collected weekly on the first three sampling days and fortnightly afterwards. The daily ambient temperatures were registered and averaged to weekly mean temperatures. Steers fed sainfoin silage had lower (P< 0.05) concentrations of branched-chain volatile fatty acids (VFA) than those fed alfalfa silage. All VFA concentrations were higher (P< 0.001) in rumen liquor samples than in fecal samples. The interaction of sample type and diet showed a significant effect (P< 0.05) on the proportions of the bacterial community that were from the phylaProteobacteriaandVerrucomicrobia.Ambient temperature had an indirect effect (P< 0.05) on the phylumFirmicutes, as it affected its proportional balance. The bacterial population diversity in samples appeared to decrease concurrently with the ambient temperature. The phylumFirmicutesexplained the first principal component at 64.83 and 42.58% of the total variance in rumen liquor and fecal samples, respectively. The sample type had a larger effect on bacterial communities than diet and temperature. Certain bacterial populations seemed to be better adapted than others to environmentally adverse conditions, such as less access time to nutrients due to higher motility and rate of passage of digesta caused by extreme temperatures, or antimicrobials such as tannins, possibly due to an influence of their biogeographical location within the gut.

scholarly journals Quantifying within- and between-animal variation and uncertainty associated with counts of Escherichia coli O157 occurring in naturally infected cattle faeces

2008 ◽  
Vol 6 (31) ◽  
pp. 169-177 ◽  
Author(s):  
S.E Robinson ◽  
P.E Brown ◽  
E.J Wright ◽  
C.A Hart ◽  
N.P French
Keyword(s):  
Escherichia Coli ◽  
Statistical Model ◽  
Human Infection ◽  
Microbial Count ◽  
Infected Cattle ◽  
Explanatory Variables ◽  
E Coli ◽  
Fixed And Random Effects ◽  
Cattle Faeces ◽  
Multiple Samples

Cattle faeces are considered the most important reservoir for human infection with Escherichia coli O157. We have previously described shedding of E. coli O157 in the faeces of naturally infected cattle cohorts. However, the data require further investigation to quantify the uncertainty and variability in the estimates previously presented. This paper proposes a method for analysing both the presence and the quantity of E. coli O157 in cattle faecal samples, using two isolation procedures, one of which enumerates E. coli O157. The combination of these two measurements, which are fundamentally different in nature and yet measuring a common outcome, has necessitated the development of a novel statistical model for ascertaining the contribution of the various components of variation (both natural and observation induced) and for judging the influence of explanatory variables. Most of the variation within the sampling hierarchy was attributable to multiple samples from the same animal. The contribution of laboratory-level variation was found to be low. After adjusting for fixed and random effects, short periods of increased intensity of shedding were identified in individual animals. We conclude that within-animal variation is greater than between animals over time, and studies aiming to elucidate the dynamics of shedding should focus resources, sampling more within than between animals. These findings have implications for the identification of persistent high shedders and for assessing their role in the epidemiology of E. coli O157 in cattle populations. The development of this non-standard statistical model may have many applications to other microbial count data.

Mannose-Binding Activity of Escherichia coli: a Determinant of Attachment and Ingestion of the Bacteria by Macrophages

1980 ◽  
Vol 29 (2) ◽  
pp. 417-424
Author(s):  
Zvi Bar-Shavit ◽  
Rachel Goldman ◽  
Itzhak Ofek ◽  
Nathan Sharon ◽  
David Mirelman
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Pathogenic Bacteria ◽  
Binding Activity ◽  
Exponential Phase ◽  
Restrictive Temperature ◽  
Filamentous Bacteria ◽  
Phagocytic Cells ◽  
E Coli ◽  
Mannose Binding

Recently, it was suggested that a mannose-specific lectin on the bacterial cell surface is responsible for the recognition by phagocytic cells of certain nonopsonized Escherichia coli strains. In this study we assessed the interaction of two strains of E. coli at different phases of growth with a monolayer of mouse peritoneal macrophages and developed a direct method with [ 14 C]mannan to quantitate the bacterial mannose-binding activity. Normal-sized bacteria were obtained from logarithmic and stationary phases of growth. Nonseptated filamentous cells were formed by growing the organisms in the presence of cephalexin or at a restrictive temperature. Attachment to macrophages of all bacterial forms was inhibited by methyl α- d -mannoside and mannan but not by other sugars tested. The attachment of stationary phase and filamentous bacteria to macrophages, as well as their mannose-binding activity, was similar, whereas in the exponential-phase bacteria they were markedly reduced. The results show a linear relation between the two parameters ( R = 0.98, P < 0.001). The internalization of the filamentous cells attached to macrophages during 45 min of incubation was much less efficient (20%) compared to that of exponential-phase, stationary-phase, or antibody-coated filamentous bacteria (90%). The results indicate that the mannose-binding activity of E. coli determines the recognition of the organisms by phagocytes. They further suggest that administration of β-lactam antibiotics may impair elimination of certain pathogenic bacteria by inducing the formation of filaments which are inefficiently internalized by the host's phagocytic cells.

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