yersiniabactin and aerobactin

The increasing prevalence of KPC-producing Klebsiella pneumoniae strains in clinical settings has been largely attributed to dissemination of organisms of specific multilocus sequence types, such as ST258 and ST11. Compared with the ST258 clone, which is prevalent in North America and Europe, ST11 is common in China but information regarding its genetic features remains scarce. In this study, we performed detailed genetic characterization of ST11 K. pneumoniae strains by analyzing whole-genome sequences of 58 clinical strains collected from diverse geographic locations in China. The ST11 genomes were found to be highly heterogeneous and clustered into at least three major lineages based on the patterns of single-nucleotide polymorphisms. Exhibiting five different capsular types, these ST11 strains were found to harbor multiple resistance and virulence determinants such as the blaKPC-2 gene, which encodes carbapenemase, and the yersiniabactin-associated virulence genes irp, ybt and fyu. Moreover, genes encoding the virulence factor aerobactin and the regulator of the mucoid phenotype (rmpA) were detectable in six genomes, whereas genes encoding salmochelin were found in three genomes. In conclusion, our data indicated that carriage of a wide range of resistance and virulence genes constitutes the underlying basis of the high level of prevalence of ST11 in clinical settings. Such findings provide insight into the development of novel strategies for prevention, diagnosis and treatment of K. pneumoniae infections.

Topics: Bacterial Proteins; beta-Lactamases; Carbapenem-Resistant Enterobacteriaceae; China; DNA, Bacterial; Drug Resistance, Multiple, Bacterial; Enterobactin; Genes, Bacterial; Genome, Bacterial; Humans; Hydroxamic Acids; Klebsiella Infections; Klebsiella pneumoniae; Microbial Sensitivity Tests; Multilocus Sequence Typing; Phenols; Polymorphism, Single Nucleotide; Sequence Alignment; Sequence Analysis; Thiazoles; Virulence; Virulence Factors

Urinary tract infections impose substantial health burdens on women worldwide. Urinary tract infections often incur a high risk of recurrence and antibiotic resistance, and uropathogenic E. coli accounts for approximately 80% of clinically acquired cases. The diagnosis of, treatment of, and drug development for urinary tract infections remain substantial challenges due to the complex pathogenesis of this condition. The clinically isolated UPEC 83972 strain was found to produce four siderophores: yersiniabactin, aerobactin, salmochelin, and enterobactin. The biosyntheses of some of these siderophores implies that the virulence of UPEC is mediated via the targeting of primary metabolism. However, the differential modulatory roles of siderophore biosyntheses on the differential metabolomes of UPEC and non-UPEC strains remain incompletely understood. In the present study, we sought to investigate how the differential metabolomes can be used to distinguish UPEC from non-UPEC strains and to determine the associated regulatory roles of siderophore biosynthesis. Our results are the first to demonstrate that the identified differential metabolomes strongly differentiated UPEC from non-UPEC strains. Furthermore, we performed metabolome assays of mutants with different patterns of siderophore deletions; the data revealed that the mutations of all four siderophores exerted a stronger modulatory role on the differential metabolomes of the UPEC and non-UPEC strains relative to the mutation of any single siderophore and that this modulatory role primarily involved amino acid metabolism, oxidative phosphorylation in the carbon fixation pathway, and purine and pyrimidine metabolism. Surprisingly, the modulatory roles were strongly dependent on the type and number of mutated siderophores. Taken together, these results demonstrated that siderophore biosynthesis coordinately modulated the differential metabolomes and thus may indicate novel targets for virulence-based diagnosis, therapeutics, and drug development related to urinary tract infections.

Topics: Amino Acids; Bacterial Proteins; Carbon Cycle; Citric Acid Cycle; Enterobactin; Escherichia coli Infections; Female; Gene Deletion; Gene Expression; Glucosides; Humans; Hydroxamic Acids; Magnetic Resonance Spectroscopy; Metabolome; Oxidative Phosphorylation; Phenols; Purines; Pyrimidines; Siderophores; Thiazoles; Urinary Tract Infections; Uropathogenic Escherichia coli; Virulence

Hypervirulent Klebsiella pneumoniae is an emerging cause of community-acquired pyogenic liver abscess. First described in Asia, it is now increasingly recognized in Western countries, commonly afflicting those with Asian descent. This raises the question of genetic predisposition versus geospecific strain acquisition. We leveraged on the Antibiotics for Klebsiella Liver Abscess Syndrome Study (A-KLASS) clinical trial ongoing in ethnically diverse Singapore, to prospectively examine the profiles of 70 patients together with their isolates' genotypic and phenotypic characteristics. The majority of isolates belonged to capsule type K1, a genetically homogenous group corresponding to sequence-type 23. The remaining K2, K5, K16, K28, K57 and K63 isolates as well as two novel cps isolates were genetically heterogeneous. K1 isolates carried higher frequencies of virulence-associated genes including rmpA (regulator of mucoid phenotype A), kfu (Klebsiella ferric uptake transporter), iuc (aerobactin), iro (salmochelin) and irp (yersiniabactin) than non-K1 isolates. The Chinese in our patient cohort, mostly non-diabetic, had higher prevalence of K1 infection than the predominantly diabetic non-Chinese (Malays, Indian and Caucasian). This differential susceptibility to different capsule types among the various ethnic groups suggests patterns of transmission (e.g. environmental source, familial transmission) and/or genetic predisposition unique to each race despite being in the same geographical location.

Topics: Anti-Bacterial Agents; Bacterial Proteins; Ethnicity; Female; Genotype; Humans; Hydroxamic Acids; Klebsiella Infections; Klebsiella pneumoniae; Liver; Male; Phenols; Serotyping; Singapore; Thiazoles; Virulence Factors

Iron is essential for Escherichia coli growth and survival in the host and the external environment, but its availability is generally low due to the poor solubility of its ferric form in aqueous environments and the presence of iron-withholding proteins in the host. Most E. coli can increase access to iron by excreting siderophores such as enterobactin, which have a very strong affinity for Fe3+. A smaller proportion of isolates can generate up to 3 additional siderophores linked with pathogenesis; aerobactin, salmochelin, and yersiniabactin. However, non-pathogenic E. coli are also able to synthesise these virulence-associated siderophores. This raises questions about their role in the ecology of E. coli, beyond virulence, and whether specific siderophores might be linked with persistence in the external environment. Under the assumption that selection favours phenotypes that confer a fitness advantage, we compared siderophore production and gene distribution in E. coli isolated either from agricultural plants or the faeces of healthy mammals. This population-level comparison has revealed that under iron limiting growth conditions plant-associated isolates produced lower amounts of siderophores than faecal isolates. Additionally, multiplex PCR showed that environmental isolates were less likely to contain loci associated with aerobactin and yersiniabactin synthesis. Although aerobactin was linked with strong siderophore excretion, a significant difference in production was still observed between plant and faecal isolates when the analysis was restricted to strains only able to synthesise enterobactin. This finding suggests that the regulatory response to iron limitation may be an important trait associated with adaptation to the non-host environment. Our findings are consistent with the hypothesis that the ability to produce multiple siderophores facilitates E. coli gut colonisation and plays an important role in E. coli commensalism.

Topics: Adaptation, Biological; Animals; Enterobactin; Environmental Microbiology; Escherichia coli; Escherichia coli Proteins; Feces; Glucosides; Humans; Hydroxamic Acids; Iron; Phenols; Plants; Siderophores; Thiazoles; Virulence Factors

The siderophore aerobactin is the dominant siderophore produced by hypervirulent Klebsiella pneumoniae (hvKP) and was previously shown to be a major virulence factor in systemic infection. However, strains of hvKP commonly produce the additional siderophores yersiniabactin, salmochelin, and enterobactin. The roles of these siderophores in hvKP infection have not been optimally defined. To that end, site-specific gene disruptions were created in hvKP1 (wild type), resulting in the generation of hvKP1ΔiucA (aerobactin deficient), hvKP1ΔiroB (salmochelin deficient), hvKP1ΔentB (enterobactin and salmochelin deficient), hvKP1Δirp2 (yersiniabactin deficient), and hvKP1ΔentBΔirp2 (enterobactin, salmochelin, and yersiniabactin deficient). The growth/survival of these constructs was compared to that of their wild-type parent hvKP1 ex vivo in human ascites fluid, human serum, and human urine and in vivo in mouse systemic infection and pulmonary challenge models. Interestingly, in contrast to aerobactin, the inability to produce enterobactin, salmochelin, or yersiniabactin individually or in combination did not decrease the ex vivo growth/survival in human ascites or serum or decrease virulence in the in vivo infection models. Surprisingly, none of the siderophores increased growth in human urine. In human ascites fluid supplemented with exogenous siderophores, siderophores increased the growth of hvKP1ΔiucA, with the relative activity being enterobactin > aerobactin > yersiniabactin > salmochelin, suggesting that the contribution of aerobactin to virulence is dependent on both innate biologic activity and quantity produced. Taken together, these data confirm and extend a role for aerobactin as a critical virulence factor for hvKP. Since it appears that aerobactin production is a defining trait of hvKP strains, this factor is a potential antivirulence target.

Topics: Animals; Enterobactin; Glucosides; Humans; Hydroxamic Acids; Klebsiella Infections; Klebsiella pneumoniae; Male; Mice; Microbial Viability; Phenols; Siderophores; Thiazoles; Virulence; Young Adult

Hypoxia inducible factor-1 (HIF-1) is the key transcriptional regulator during adaptation to hypoxia. Recent studies provide evidence for HIF-1 activation during bacterial infections. However, molecular details of how bacteria activate HIF-1 remain unclear. Here, we pursued the role of bacterial siderophores in HIF-1 activation during infection with Enterobacteriaceae.. In vivo, HIF-1 activation and HIF-1-dependent gene induction in Peyer's patches were analyzed after orogastric infection with Yersinia enterocolitica. The course of an orogastric Y enterocolitica infection was determined using mice with a deletion of HIF-1alpha in the intestine. In vitro, the mechanism of HIF-1 activation was analyzed in infections with Y enterocolitica, Salmonella enterica subsp enterica, and Enterobacter aerogenes.. Infection of mice with Y enterocolitica led to functional activation of HIF-1 in Peyer's patches. Because mice with deletion of HIF-1alpha in the intestinal epithelium showed a significantly higher susceptibility to orogastric Y enterocolitica infections, bacterial HIF-1 activation appears to represent a host defense mechanism. Additional studies with Y enterocolitica, S enterica subsp enterica, or E aerogenes, and, moreover, application of their siderophores (yersiniabactin, salmochelin, aerobactin) caused a robust, dose-dependent HIF-1 response in human epithelia and endothelia, independent of cellular hypoxia. HIF-1 activation occurs most likely because of inhibition of prolylhydroxylase activity and is abolished upon infection with siderophore uptake deficient bacteria.. Taken together, this study reveals what we believe to be a previously unrecognized role of bacterial siderophores for hypoxia-independent activation of HIF-1 during infection with human pathogenic bacteria.

Topics: Animals; Caco-2 Cells; Cell Hypoxia; Disease Models, Animal; Endothelial Cells; Enterobacter aerogenes; Enterobacteriaceae; Epithelial Cells; Female; Gene Expression Regulation; HeLa Cells; Humans; Hydroxamic Acids; Hydroxylation; Hypoxia-Inducible Factor 1, alpha Subunit; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Oxygen; Peyer's Patches; Phenols; Procollagen-Proline Dioxygenase; Salmonella enterica; Siderophores; Thiazoles; Time Factors; Transcriptional Activation; Up-Regulation; Yersinia enterocolitica; Yersinia Infections

Escherichia coli is the most frequent microorganism involved in urinary tract infection (UTI). Acute UTI caused by uropathogenic E. coli (UPEC) can lead to recurrent infection, which can be defined as either re-infection or relapse. E. coli strains causing relapse (n = 27) and re-infection (n = 53) were analysed. In-vitro production of biofilm, yersiniabactin and aerobactin was significantly more frequent among strains causing relapse. Biofilm assays may be helpful in selecting patients who require a therapeutic approach to eradicate persistent biofilm-forming E. coli strains and prevent subsequent relapses.

Topics: Adult; Aged; Aged, 80 and over; Biofilms; Escherichia coli; Escherichia coli Infections; Female; Humans; Hydroxamic Acids; Middle Aged; Phenols; Thiazoles; Urinary Tract Infections; Virulence

The probiotic Escherichia coli strain Nissle 1917 produces four siderophores: the catecholates enterobactin and salmochelin, the hydroxamate aerobactin, and the mixed-type siderophore yersiniabactin. We studied the influence of pH, temperature, and carbon source on the production of these four siderophores. Yersiniabactin and salmochelin were maximally produced under neutral to alkaline conditions (pH 7.0 and 7.6, respectively), whereas aerobactin was maximally produced at a more acidic pH (pH 5.6), which agrees with the slightly higher complex stability of hydroxamates at acidic pH values compared to the catecholates. Under nearly all conditions studied, catecholate siderophore production was higher with glycerol than with glucose as the carbon source. Yersiniabactin production was also higher with glycerol as the carbon source at pH 7.0. At 42 degrees C, strain Nissle 1917 grew poorly or not at all because of the iron-limiting conditions. In a competition experiment between wild-type strain Nissle 1917 and a mutant of this strain with a deletion in the yersiniabactin operon, the wild-type overgrew the mutant at pH 7.0 and 7.6 and not at pH 5.6. These results agree with yersiniabactin production being of greater advantage at neutral and slightly alkaline pH values. The production of four siderophores may help the probiotic E. coli Nissle 1917 to compete with other E. coli strains in the colon. The probiotic strain Nissle 1917 used in our experiments has many characteristics in common with uropathogenic E. coli and other pathogenic strains which also secrete these siderophores. Uropathogenic E. coli strains may need the multitude of siderophores to adapt to the pH of urine, which varies between pH 4.6 and 8.0.

Topics: Chromatography, High Pressure Liquid; Enterobactin; Environment; Escherichia coli; Ferric Compounds; Glucosides; Hydrogen-Ion Concentration; Hydroxamic Acids; Phenols; Siderophores; Thiazoles

We analyzed the ability of extraintestinal strains of Enterobacter spp. and Citrobacter spp. to employ different siderophore-mediated strategies of iron acquisition. All strains produced iron-chelating compounds. Cross-feeding assays indicated that most isolates of both Enterobacter spp. and Citrobacter spp. excreted catecholate siderophore enterobactin, less produced aerobactin, and single strains excreted hydroxamates different from aerobactin. Besides, we analyzed if the strains had the ability to produce the siderophore yersiniabactin coded by the Yersinia high-pathogenicity island (HPI). The presence of HPI genes was observed in single isolates of three species: E. cloaceae, E. aerogenes and C. koseri. A detailed polymerase chain reaction analysis revealed differences in the genetic organization of the HPIs; however, in a cross-feeding test we proved that yersiniabactin was produced and the island was functional.

Topics: Bacterial Proteins; Chelating Agents; Citrobacter; DNA, Bacterial; Enterobacter; Enterobacteriaceae Infections; Genomic Islands; Humans; Hydroxamic Acids; Iron; Phenols; Polymerase Chain Reaction; Siderophores; Thiazoles; Virulence; Yersinia

The ability to acquire iron is crucial for bacteria during an infection. The capacity of 35 strains of Escherichia coli, isolated from clinical specimens, to use various strategies to obtain iron was analysed. The isolates employed several iron-uptake mechanisms, including production of enterobactin (86 %) and aerobactin (71 %). The majority of the isolates also excreted yersiniabactin, which is encoded by the Yersinia high-pathogenicity island (HPI). However, PCR analysis of the Yersinia HPI revealed diversity in its genetic organization. Use of human transferrin (91 %), lactoferrin (94 %), haemoglobin (80 %) and haemoglobin-haptoglobin complex (63 %) as the sole source of iron was common among E. coli isolates. Multiple iron-uptake systems may be of benefit to bacteria during an infection.

Topics: Bacterial Outer Membrane Proteins; Bacterial Proteins; Enterobactin; Escherichia coli; Genes, Bacterial; Hemolysis; Humans; Hydroxamic Acids; Iron; Iron-Binding Proteins; Multigene Family; Periplasmic Binding Proteins; Phenols; Siderophores; Thiazoles; Yersinia

The ability to acquire iron is crucial to bacteria during an infection. Thirty-four strains of Klebsiella pneumoniae isolated from clinical specimens were examined for the use of various strategies to obtain iron. The isolates employed several iron uptake mechanisms, including production of enterobactin (100%) and aerobactin (50%). Few isolates (18%) produced yersiniabactin, a siderophore encoded by the Yersinia high-pathogenicity island (HPI) despite genetic diversity of the HPI. Majority of the isolates used human transferrin (74%), lactoferrin (97%), hemoglobin (74%), and hemoglobin-haptoglobin complex (56%) as a sole source of iron. Multiple iron uptake systems may be of benefit to the bacteria during infection.

Topics: Chromosomes, Bacterial; DNA, Bacterial; Enterobactin; Gene Order; Genomic Islands; Haptoglobins; Hemoglobins; Hemolysin Proteins; Humans; Hydroxamic Acids; Iron; Klebsiella pneumoniae; Lactoferrin; Phenols; Polymerase Chain Reaction; Siderophores; Thiazoles; Transferrin; Yersinia

High pathogenicity islands (HPIs), first identified in various Yersinia species, encode an iron uptake system. We have studied the occurrence of HPIs in septicemic strains of Escherichia coli isolated from a variety of hosts. The results presented in this communication indicate that most septicemic strains tested contained HPI sequences even though they already have the aerobactin encoding genes. We have also observed two types of HPI deletions, suggesting genetic instability of this element. Notable exceptions are several strains isolated from septicemia in sheep that lacked both iron acquisition systems.

Topics: Animals; Bacteremia; Bacterial Outer Membrane Proteins; Bacterial Proteins; Cattle; Cattle Diseases; Escherichia coli; Escherichia coli Infections; Genes, Bacterial; Humans; Hydroxamic Acids; Infant, Newborn; Iron-Binding Proteins; Meningitis, Escherichia coli; Mixed Function Oxygenases; Periplasmic Binding Proteins; Phenols; Polymerase Chain Reaction; Poultry; Poultry Diseases; Receptors, Cell Surface; Serotyping; Sheep; Sheep Diseases; Siderophores; Thiazoles; Virulence