scholarly journals Role of Glucose in Enhancing the Temperature-Dependent Growth Inhibition of Escherichia coli O157:H7 ATCC 43895 by a Pseudomonas sp

2002 ◽  
Vol 68 (5) ◽  
pp. 2600-2604 ◽  
Author(s):  
John Samelis ◽  
John N. Sofos

ABSTRACT Growth of Escherichia coli O157:H7 strain ATCC 43895 was monitored at 5, 10, 15, and 25°C in both pure and mixed (1:1) cultures with a gluconate-producing Pseudomonas sp. found in meat to evaluate the effect of the absence and presence of 1% glucose in broth on temperature-dependent competition. The number of colonies of the Pseudomonas strain exceeded 9 log CFU/ml under all conditions tested. The pathogen grew better as the temperature increased from 10 to 15 and 25°C and grew better in pure culture than in mixed cultures. Pseudomonas sp. inhibited E. coli O157:H7 in cocultures with glucose at 10°C, while at 15°C the pathogen exhibited a biphasic pattern of growth with an intermediate inactivation period. Pathogen inhibition was much weaker in cocultures grown without glucose at 10 to 15°C and, irrespective of glucose, at 25°C. These results indicate that glucose enhances the growth inhibition of E. coli O157:H7 by some Pseudomonas spp., potentially due to its rapid uptake and conversion to gluconate, at low (≤15°C) temperatures.

mBio ◽  
2013 ◽  
Vol 4 (4) ◽  
Author(s):  
Zachary C. Ruhe ◽  
Adam B. Wallace ◽  
David A. Low ◽  
Christopher S. Hayes

ABSTRACT Bacteria that express contact-dependent growth inhibition (CDI) systems outcompete siblings that lack immunity, suggesting that CDI mediates intercellular competition. To further explore the role of CDI in competition, we determined the target cell range of the CDIEC93 system from Escherichia coli EC93. The CdiAEC93 effector protein recognizes the widely conserved BamA protein as a receptor, yet E. coli EC93 does not inhibit other enterobacterial species. The predicted membrane topology of BamA indicates that three of its extracellular loops vary considerably between species, suggesting that loop heterogeneity may control CDI specificity. Consistent with this hypothesis, other enterobacteria are sensitized to CDIEC93 upon the expression of E. coli bamA and E. coli cells become CDIEC93 resistant when bamA is replaced with alleles from other species. Our data indicate that BamA loops 6 and 7 form the CdiAEC93-binding epitope and their variation between species restricts CDIEC93 target cell selection. Although BamA loops 6 and 7 vary dramatically between species, these regions are identical in hundreds of E. coli strains, suggesting that BamAEcoli and CdiAEC93 play a role in self-nonself discrimination. IMPORTANCE Contact-dependent growth inhibition (CDI) systems are widespread among Gram-negative bacteria, enabling them to bind to neighboring bacterial cells and deliver protein toxins that inhibit cell growth. In this study, we tested the role of CDI in interspecies competition using intestinal isolate Escherichia coli EC93 as an inhibitor cell model. Although E. coli EC93 inhibits different E. coli strains, other bacterial species from the intestine are completely resistant to CDI. We show that resistance is due to small variations in the CDI receptor that prevent other species from being recognized as target cells. CDI receptor interactions thus provide a mechanism by which bacteria can distinguish siblings and other close relatives (self) from more distant relatives or other species of bacteria (nonself). Our results provide a possible means by which antimicrobials could be directed to one or only a few related bacterial pathogens by using a specific receptor “zip code.”


2005 ◽  
Vol 68 (12) ◽  
pp. 2559-2566 ◽  
Author(s):  
SYLVIA GAYSINSKY ◽  
P. MICHAEL DAVIDSON ◽  
BARRY D. BRUCE ◽  
JOCHEN WEISS

Growth inhibition of four strains of Escherichia coli O157:H7 (H1730, F4546, 932, and E0019) and Listeria monocytogenes (Scott A, 101, 108, and 310) by essential oil components (carvacrol and eugenol) solubilized in nonionic surfactant micelles (Surfynol 465 and 485W) was investigated. Concentrations of encapsulated essential oil components ranged from 0.02 to 1.25% depending on compound, surfactant type, and surfactant concentration (0.5 to 5%). Eugenol encapsulated in Surfynol 485W micelles was most efficient in inhibiting growth of the pathogens; 1% Surfynol 485W and 0.15% eugenol was sufficient to inhibit growth of all strains of E. coli O157:H7 and three of four strains of L. monocytogenes (Scott A, 310, and 108). The fourth strain, L. monocytogenes 101, was inhibited by 2.5% Surfynol and 0.225% eugenol. One percent Surfynol 485W in combination with 0.025% carvacrol was effective in inhibiting three of four strains of E. coli O157:H7. Strain H1730 was the most resistant strain, requiring 0.3% carvacrol and 5% surfactant for complete inhibition. Growth inhibition of L. monocytogenes by combinations of carvacrol and Surfynol 465 ranged between 0.15 and 0.35% and 1 and 3.75%, respectively. Generally, the antimicrobial activity of Surfynol 465 in combination with eugenol was higher than that for the combination with carvacrol. The potent activity was attributed to increased solubility of essential oil components in the aqueous phase due to the presence of surfactants and improved interactions of antimicrobials with microorganisms.


2001 ◽  
Vol 64 (2) ◽  
pp. 147-151 ◽  
Author(s):  
KAZUE TAKEUCHI ◽  
JOSEPH F. FRANK

Viability of Escherichia coli O157:H7 cells on lettuce leaves after 200 mg/liter (200 ppm) chlorine treatment and the role of lettuce leaf structures in protecting cells from chlorine inactivation were evaluated by confocal scanning microscopy (CSLM). Lettuce samples (2 by 2 cm) were inoculated by immersing in a suspension containing 109 CFU/ml of E. coli O157: H7 for 24 ± 1 h at 4°C. Rinsed samples were treated with 200 mg/liter (200 ppm) chlorine for 5 min at 22°C. Viability of E. coli O157:H7 cells was evaluated by CSLM observation of samples stained with Sytox green (dead cell stain) and Alexa 594 conjugated antibody against E. coli O157:H7. Quantitative microscopic observations of viability were made at intact leaf surface, stomata, and damaged tissue. Most E. coli O157:H7 cells (68.3 ± 16.2%) that had penetrated 30 to 40 μm from the damaged tissue surface remained viable after chlorine treatment. Cells on the surface survived least (25.2 ± 15.8% survival), while cells that penetrated 0 to 10 μm from the damaged tissue surface or entered stomata showed intermediate survival (50.8 ± 13.5 and 45.6 ± 9.7% survival, respectively). Viability was associated with the depth at which E. coli O157:H7 cells were in the stomata. Although cells on the leaf surface were mostly inactivated, some viable cells were observed in cracks of cuticle and on the trichome. These results demonstrate the importance of lettuce leaf structures in the protection of E. coli O157:H7 cells from chlorine inactivation.


2015 ◽  
Vol 12 (110) ◽  
pp. 20150446 ◽  
Author(s):  
Simon E. F. Spencer ◽  
Thomas E. Besser ◽  
Rowland N. Cobbold ◽  
Nigel P. French

Supershedders have been suggested to be major drivers of transmission of Escherichia coli O157:H7 ( E. coli O157:H7) among cattle in feedlot environments, despite our relatively limited knowledge of the processes that govern periods of high shedding within an individual animal. In this study, we attempt a data-driven approach, estimating the key characteristics of high shedding behaviour, including effects on transmission to other animals, directly from a study of natural E. coli O157:H7 infection of cattle in a research feedlot, in order to develop an evidence-based definition of supershedding. In contrast to the hypothesized role of supershedders, we found that high shedding individuals only modestly increased the risk of transmission: individuals shedding over 10 3 cfu g −1 faeces were estimated to pose a risk of transmission only 2.45 times greater than those shedding below that level. The data suggested that shedding above 10 3 cfu g −1 faeces was the most appropriate definition of supershedding behaviour and under this definition supershedding was surprisingly common, with an estimated prevalence of 31.3% in colonized individuals. We found no evidence that environmental contamination by faeces of shedding cattle contributed to transmission over timescales longer than 3 days and preliminary evidence that higher stocking density increased the risk of transmission.


2009 ◽  
Vol 75 (20) ◽  
pp. 6515-6523 ◽  
Author(s):  
Terrance M. Arthur ◽  
James E. Keen ◽  
Joseph M. Bosilevac ◽  
Dayna M. Brichta-Harhay ◽  
Norasak Kalchayanand ◽  
...  

ABSTRACT The objectives of the study described here were (i) to investigate the dynamics of Escherichia coli O157:H7 fecal and hide prevalence over a 9-month period in a feedlot setting and (ii) to determine how animals shedding E. coli O157:H7 at high levels affect the prevalence and levels of E. coli O157:H7 on the hides of other animals in the same pen. Cattle (n = 319) were distributed in 10 adjacent pens, and fecal and hide levels of E. coli O157:H7 were monitored. When the fecal pen prevalence exceeded 20%, the hide pen prevalence was usually (25 of 27 pens) greater than 80%. Sixteen of 19 (84.2%) supershedder (>104 CFU/g) pens had a fecal prevalence greater than 20%. Significant associations with hide and high-level hide (≥40 CFU/100 cm2) contamination were identified for (i) a fecal prevalence greater than 20%, (ii) the presence of one or more high-density shedders (≥200 CFU/g) in a pen, and (iii) the presence of one or more supershedders in a pen. The results presented here suggest that the E. coli O157:H7 fecal prevalence should be reduced below 20% and the levels of shedding should be kept below 200 CFU/g to minimize the contamination of cattle hides. Also, large and unpredictable fluctuations within and between pens in both fecal and hide prevalence of E. coli O157:H7 were detected and should be used as a guide when preharvest studies, particularly preharvest intervention studies, are designed.


2006 ◽  
Vol 188 (21) ◽  
pp. 7661-7667 ◽  
Author(s):  
Ahmed Touhami ◽  
Manfred Jericho ◽  
Andrew D. Rutenberg

ABSTRACT We observed that the oscillation period of MinD within rod-like and filamentous cells of Escherichia coli varied by a factor of 4 in the temperature range from 20°C to 40°C. The detailed dependence was Arrhenius, with a slope similar to the overall temperature-dependent growth curve of E. coli. The detailed pattern of oscillation, including the characteristic wavelength in filamentous cells, remained independent of temperature. A quantitative model of MinDE oscillation exhibited similar behavior, with an activated temperature dependence of the MinE-stimulated MinD-ATPase rate.


2005 ◽  
Vol 71 (1) ◽  
pp. 247-254 ◽  
Author(s):  
Jee-Hoon Ryu ◽  
Larry R. Beuchat

ABSTRACT The resistance of Escherichia coli O157:H7 strains ATCC 43895-, 43895-EPS (an exopolysaccharide [EPS]-overproducing mutant), and ATCC 43895+ (a curli-producing mutant) to chlorine, a sanitizer commonly used in the food industry, was studied. Planktonic cells of strains 43895-EPS and/or ATCC 43895+ grown under conditions supporting EPS and curli production, respectively, showed the highest resistance to chlorine, indicating that EPS and curli afford protection. Planktonic cells (ca. 9 log10 CFU/ml) of all strains, however, were killed within 10 min by treatment with 50 μg of chlorine/ml. Significantly lower numbers of strain 43895-EPS, compared to those of strain ATCC 43895-, attached to stainless steel coupons, but the growth rate of strain 43895-EPS on coupons was not significantly different from that of strain ATCC 43895-, indicating that EPS production did not affect cell growth during biofilm formation. Curli production did not affect the initial attachment of cells to coupons but did enhance biofilm production. The resistance of E. coli O157:H7 to chlorine increased significantly as cells formed biofilm on coupons; strain ATCC 43895+ was the most resistant. Population sizes of strains ATCC 43895+ and ATCC 43895- in biofilm formed at 12�C were not significantly different, but cells of strain ATCC 43895+ showed significantly higher resistance than did cells of strain ATCC 43895-. These observations support the hypothesis that the production of EPS and curli increase the resistance of E. coli O157:H7 to chlorine.


2008 ◽  
Vol 76 (11) ◽  
pp. 5062-5071 ◽  
Author(s):  
Alfredo G. Torres ◽  
Terry M. Slater ◽  
Shilpa D. Patel ◽  
Vsevolod L. Popov ◽  
Margarita M. P. Arenas-Hernández

ABSTRACT The expression of the long polar fimbriae (LPF) of enterohemorrhagic Escherichia coli (EHEC) O157:H7 is controlled by a tightly regulated process, and, therefore, the role of these fimbriae during binding to epithelial cells has been difficult to establish. We recently found that histone-like nucleoid-structuring protein (H-NS) binds to the regulatory sequence of the E. coli O157:H7 lpf1 operon and “silences” its transcription, while Ler inhibits the action of the H-NS protein and allows lpf1 to be expressed. In the present study, we determined how the deregulated expression of LPF affects binding of EHEC O157:H7 to tissue-cultured cells, correlating the adherence phenotype with lpf1 expression. We tested the adherence properties of EHEC hns mutant and found that this strain adhered 2.8-fold better than the wild type. In contrast, the EHEC ler mutant adhered 2.1-fold less than the wild type. The EHEC hns ler mutant constitutively expressed the lpf genes, and, therefore, we observed that the double mutant adhered 5.6-fold times better than the wild type. Disruption of lpfA in the EHEC hns and hns ler mutants or the addition of anti-LpfA serum caused a reduction in adhesion, demonstrating that the increased adherence was due to the expression of LPF. Immunogold-labeling electron microscopy showed that LPF is present on the surface of EHEC lpfA + strains. Furthermore, we showed that EHEC expressing LPF agglutinates red blood cells from different species and that the agglutination was blocked by the addition of anti-LpfA serum. Overall, our data confirmed that expression of LPF is a tightly regulated process and, for the first time, demonstrated that these fimbriae are associated with adherence and hemagglutination phenotypes in EHEC O157:H7.


2020 ◽  
Author(s):  
Clément Patacq ◽  
Nicolas Chaudet ◽  
Fabien Letisse

ABSTRACTBacteria grow in constantly changing environments that can suddenly become completely deleted in essential nutrients. The stringent response, a rewiring of the cellular metabolism mediated by the alarmone (p)ppGpp, plays a crucial role in adjusting bacterial growth to the severity of the nutritional insult. The ability of (p)ppGpp to trigger a slowdown of cell growth or induce bacterial dormancy has been widely investigated. However, little is known about the role of (p)ppGpp in promoting growth recovery after severe growth inhibition. In this study, we performed a time-resolved analysis of (p)ppGpp metabolism in Escherichia coli as it recovered from a sudden slowdown in growth. Results show that E. coli recovers by itself from the growth disruption provoked by the addition of serine hydroxamate, the serine analogue that we used to induce the stringent response. Growth inhibition was accompanied by a severe disturbance of metabolic activity and more surprisingly, by a transient overflow of valine and alanine. Our data also show that ppGpp is crucial for growth recovery since in the absence of ppGpp, E. coli’s growth recovery was slower. In contrast, an increased concentration of pppGpp was found to have no significant effect on growth recovery. Interestingly, the observed decrease in intracellular ppGpp levels in the recovery phase correlated with bacterial growth and the main effect involved was identified as growth dilution rather than active degradative process. This report thus significantly expands our knowledge of (p)ppGpp metabolism in E. coli physiology.IMPORTANCEThe capacity of microbes to resist and overcome environmental insults, know as resilience, allows them to survive in changing environments but also to resist antibiotic and biocide treatments, immune system responses. Although the role of the stringent response in bacterial resilience to nutritional insults has been well studied, little is known about its importance in the ability of the bacteria to not just resist but also recover from these disturbances. To address this important question, we investigated growth disruption resilience in the model bacterium Escherichia coli and its dependency on the stringent response alarmone (p)ppGpp by quantifying ppGpp and pppGpp levels as growth was disrupted and then recovered. Our findings may thus contribute to understanding how ppGpp improves E. coli’s resilience to nutritional stress and other environmental insults.


2005 ◽  
Vol 73 (3) ◽  
pp. 1836-1846 ◽  
Author(s):  
Angus Best ◽  
Roberto M. La Ragione ◽  
A. Robin Sayers ◽  
Martin J. Woodward

ABSTRACT Shiga toxin (Stx)-positive Escherichia coli O157:H7 readily colonize and persist in specific-pathogen-free (SPF) chicks, and we have shown that an Stx-negative E. coli O157:H7 isolate (NCTC12900) readily colonizes SPF chicks for up to 169 days after oral inoculation at 1 day of age. However, the role of intimin in the persistent colonization of poultry remains unclear. Thus, to investigate the role of intimin and flagella, which is a known factor in the persistence of non-O157 E. coli in poultry, isogenic single- and double-intimin and aflagellar mutants were constructed in E. coli O157:H7 isolate NCTC12900. These mutants were used to inoculate (105 CFU) 1-day-old SPF chicks. In general, significant attenuation of the aflagellate and intimin-aflagellate mutants, but not the intimin mutant, was noted at similar time points between 22 and 92 days after inoculation. The intimin-deficient mutant was still being shed at the end of the experiment, which was 211 days after inoculation, 84 days more than the wild type. Shedding of the aflagellar and intimin-aflagellar mutants ceased 99 and 113 days after inoculation, respectively. Histological analysis of gastrointestinal tissues from inoculated birds gave no evidence for true microcolony formation by NCTC12900 or intimin and aflagellar mutants to epithelial cells. However, NCTC12900 mutant derivatives associated with the mucosa were observed as individual cells and/or as large aggregates. Association with luminal contents was also noted. These data suggest that O157 organisms do not require intimin for the persistent colonization of chickens, whereas flagella do play a role in this process.


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