A simple artificial urine for the growth of urinary pathogens

Brooks, Timothy; Keevil, C. W.

doi:10.1046/j.1472-765x.1997.00378.x

Cited by 458 publications

(379 citation statements)

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“…coli CFT073 grows more rapidly on acetogenic substrates than E. coli MG1655. Urine is analogous to tryptone broth insofar as both are poor sources of free carbohydrate but contain high levels of peptides and amino acids (8). When growing in tryptone broth, E. coli bacteria consume amino acids in a specific order, starting with serine, and favor production of acetate during early-log-phase growth (1).…”

Section: Resultsmentioning

confidence: 99%

“…The host defense involves phagocytic attack, antimicrobial peptides, complement lytic and opsonizing factors, and reactive oxygen and nitrogen species. In addition, the urinary tract as reflected in urine is limited in nutrients common to the intestinal tract, especially neutral sugars and iron (8). We hypothesize that UPEC adapted to take advantage of nutrients present in urine to more effectively colonize the urinary tract than the typical fecal E. coli strains, which rarely cause symptomatic UTIs.…”

mentioning

confidence: 99%

“…From the standpoint of E. coli nutrition, urine is a dilute mixture of amino acids and small peptides, quite similar to tryptone broth with the notable exception of the abundance of urea in urine, a molecule that E. coli cannot utilize (8). The growth of E. coli in tryptone broth cultures is well characterized.…”

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confidence: 99%

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Uropathogenic Escherichia coli CFT073 Is Adapted to Acetatogenic Growth but Does Not Require Acetate during Murine Urinary Tract Infection

et al. 2008

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In vivo accumulation of D-serine by Escherichia coli CFT073 leads to elevated expression of PAP fimbriae and hemolysin by an unknown mechanism. Loss of D-serine catabolism by CFT073 leads to a competitive advantage during murine urinary tract infection (UTI), but loss of both D-and L-serine catabolism results in attenuation. Serine is the first amino acid to be consumed in closed tryptone broth cultures and precedes the production of acetyl phosphate, a high-energy molecule involved in intracellular signaling, and the eventual secretion of acetate. We propose that the colonization defect associated with the loss of serine catabolism is due to perturbations of acetate metabolism. CFT073 grows more rapidly on acetogenic substrates than does E. coli K-12 isolate MG1655. As shown by transcription microarray results, D-serine is catabolized into acetate via the phosphotransacetylase (pta) and acetate kinase (ackA) genes while downregulating expression of acetyl coenzyme A synthase (acs). CFT073 acs, which is unable to reclaim secreted acetate, colonized mouse bladders and kidneys in the murine model of UTI indistinguishably from the wild type. Both pta and ackA are involved in the maintenance of intracellular acetyl phosphate. CFT073 pta and ackA mutants were screened to investigate the role of acetyl phosphate in UTI pathogenesis. Both single mutants are at a competitive disadvantage relative to the wild type in the kidneys but normally colonize the bladder. CFT073 ackA pta was attenuated in both the bladder and the kidneys. Thus, we demonstrate that CFT073 is adapted to acetate metabolism as a result of requiring a proper cycling of the acetyl phosphate pathway for colonization of the upper urinary tract.Urinary tract infections (UTIs) place a significant burden on the United States healthcare system, costing upwards of $2.4 billion per year (28). The majority of women will experience a UTI in their lifetime, and every year there are approximately 6.8 million physician visits, 1.2 million emergency room visits, a quarter million hospitalizations, and thousands of deaths due to complications of UTIs, most often sepsis (10,18,28,47). Escherichia coli is the most commonly isolated causative agent of community-acquired UTIs (10).The transition from residence in the gastrointestinal tract, where uropathogenic Escherichia coli (UPEC) transiently resides, to the urinary tract represents a significant change in environment. While the gastrointestinal tract is densely populated with many different species of bacteria, the bladder is normally a sterile environment yet one that presents significant challenges to bacterial growth. In addition to the cleansing flow of urine, numerous innate and acquired immune factors challenge the growth of UPEC in the urinary tract. The host defense involves phagocytic attack, antimicrobial peptides, complement lytic and opsonizing factors, and reactive oxygen and nitrogen species. In addition, the urinary tract as reflected in urine is limited in nutrients common to the intestinal tract, esp...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Uropathogenic Escherichia coli CFT073 Is Adapted to Acetatogenic Growth but Does Not Require Acetate during Murine Urinary Tract Infection

et al. 2008

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“…An important step in such a standardization is the use of a reproducible growth medium dedicated to UTP. Artificial urine formulations might be such a valuable medium, since they have a reproducible composition and allow growth of the most common UTP [11]. In this study, we combined the use of artificial urine with microcalorimetry to improve and standardize detection of UTP by IMC.…”

Section: Introductionmentioning

confidence: 99%

Standardization of isothermal microcalorimetry in urinary tract infection detection by using artificial urine

et al. 2012

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“…31 Supplementary Table 1 demonstrates molarities of each salt used for the preparation of AU. The appropriate amounts of NH3 were added from a 30% ammonium hydroxide stock solution.…”

Section: Preparation Of Artificial Urinementioning

confidence: 99%

Simple, Robust, and Plasticizer-Free Iodide-Selective Sensor Based on Copolymerized Triazole-Based Ionic Liquid

Mendecki

Chen

Callan³

et al. 2016

Anal. Chem.

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Novel solid contact iodide selective electrodes based on covalently attached 1,2,3 triazole ionic liquid (IL) were prepared and investigated in this study. Triazole-based IL moieties were synthesized using click chemistry and were further copolymerized with lauryl methacrylate via a simple one step free radical polymerization to produce a "self-plasticized" copolymer. The mechanical properties of the copolymer are suitable for the fabrication of plasticizer-free ion-selective membrane electrodes. We demonstrate that covalently attached IL moieties provide adequate functionality to the ion selective membrane thus achieving a very simple, one component sensing membrane. We also demonstrate the presence of iodide as the counter-ion in the triazole moiety has direct influence on membrane's functionality. Potentiometric experiments revealed that each electrode displays high selectivity towards iodide anions over a number of inorganic anions. Moreover the inherent presence of the iodide in the membrane reduces the need for conditioning. The non-conditioned electrodes show strikingly similar response characteristics compared to the conditioned ones. The electrodes exhibited a near Nernstian behavior with a slope of -56.1 mV per decade across large concentration range with lower detection limits found at approximately 6.3x10 -8 M or 8 ppb . These all-solid state sensors were utilized for the selective potentiometric determination of iodide ions in artificial urine samples in the nanomolar concentration range.Potentiometric chemical sensors, with primary responses based on extraction and molecular recognition processes are a wellstudied and understood class of sensing devices. 1 Ion selective electrodes (ISEs) have been already widely used in a variety of fields such as clinical analysis, 2 process control 3 and environmental monitoring. 4 Ion selective membranes are typically composed of plasticized polymers, ion exchange salts, and one or more ionophores. Each constituent plays a specific role in the proper functioning of these membrane based ISEs. 5 Spontaneous and non-specific extraction of analyte ions from the sample into the membrane bulk is primarily suppressed due to the highly hydrophobic nature of the polymer backbone. Ideally, polymer matrix should provide a homogenous medium in which all active components can move freely. This strongly resembles the composition of liquid membrane electrodes since their sensing components were simply dissolved in an organic medium. However, the performance of polymer-based membranes can be drastically reduced if such sensors are used for the measurements of ions within more lipophilic environments in biological samples including undiluted whole blood. The cross contamination of chemical sensors coupled with leaching of the sensing components from the ion selective membrane into the sample fundamentally limited the applications of ISEs as a robust analytical tool for long-term trace level analysis. 6 Over the years, a number of approaches have been developed to minim...

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A simple artificial urine for the growth of urinary pathogens

Cited by 458 publications

References 0 publications

Uropathogenic Escherichia coli CFT073 Is Adapted to Acetatogenic Growth but Does Not Require Acetate during Murine Urinary Tract Infection

Uropathogenic Escherichia coli CFT073 Is Adapted to Acetatogenic Growth but Does Not Require Acetate during Murine Urinary Tract Infection

Standardization of isothermal microcalorimetry in urinary tract infection detection by using artificial urine

Simple, Robust, and Plasticizer-Free Iodide-Selective Sensor Based on Copolymerized Triazole-Based Ionic Liquid

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