Towards discrimination of mammary pathogenic Escherichia coli (MPEC) in cattle based on possession of different iron acquisition systems

AbstractMost efforts to elucidate virulence mechanisms of mammary pathogenic Escherichia coli (MPEC), causative agent of bovine clinical mastitis, have been failed but some recent studies introduced iron acquisition systems as major role players in pathogenicity. Here, we investigated the different iron uptake systems genotypes and assessed how they relate to virulence potential of MPEC. In total, 217 E. coli isolates (MPEC= 157, fecal isolates= 60) were screened for the presence of nine genes related to iron acquisition (iroN, iutA, fecA, fyuA, sitA, irp2, iucD, chuA and tonB) and phylogenetic groups were also determined. Next, bacterial growth potential and survival in raw and UHT milk which are representative for crucial steps in mastitis development were evaluated. In addition, the mineral consumption of E. coli cultured in milk were measured. The results showed that MPEC strains considerably tend to possess fecA (93%, p= 0.000) and belong to phylogenetic group A (42%, p= 0.042). The fecA+ strains from both mastitis and fecal E. coli had a significant (p= 0.000) growth potential in raw and UHT milk. Interestingly, for the first time, it was shown that fecA+ isolates consumed less amounts of iron and other metal ions. Overall, it seems that the uptake systems related to fecA contributes to overcoming harsh conditions of milk and genetic lineages could also affect pathogenicity of MPEC. These findings could lead us to define MPEC with more clarity based on genotypes or growth potential in milk and possibly promote novel solutions to control mastitis more effectively in future.ImportanceMastitis is one of the most costly concerns in bovine medicine and the main cause of antibiotic use in dairy herds’ worldwide. As a rule of thumb, it was believed that extraintestinal pathogenic E. coli expands iron acquisition virulence arsenal to enhance pathogenic potential in the environments of the host outside the intestines. The present study indicated that the long believed idea of possession of diverse mineral acquisition systems and siderophores in all ExPEC groups could be a fairy tale. Along with recent studies, the present research showed that the fec operon could be the minimal necessary factor to overcome the harsh conditions of milk with limiting mineral concentrations. Obviously, the fecA+ isolates were fast-growing and consumed less amounts of minerals. It seems that the fec locus and its related metabolic pathways could be the potential targets for diagnostic, preventive, and therapeutic purposes.

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Allelic types of long polar fimbriae in bovine and human Escherichia coli O157 strains

Acta Veterinaria Hungarica ◽

10.1556/avet.2012.001 ◽

2012 ◽

Vol 60 (1) ◽

pp. 1-15 ◽

Cited By ~ 4

Author(s):

Domonkos Sváb ◽

István Tóth

Keyword(s):

Escherichia Coli ◽

Escherichia Coli O157 ◽

Phylogenetic Groups ◽

Genetic Lineages ◽

E Coli ◽

Typing Scheme ◽

Healthy Cattle ◽

Reference Collection ◽

Group D ◽

Atypical Strains

Long polar fimbriae (Lpf) are recently discovered adhesins and increasingly important genetic markers of pathogenicEscherichia colistrains. The presence and genotype diversity of Lpf operons was screened in a collection of 97Escherichia coliO157 strains representing different pathotypes, isolated from healthy cattle (n = 43) and human patients (n = 54) in several countries. Individual structural genes of Lpf were scanned by PCR, and allelic variants were detected with a recently developed typing scheme. Ninety-five strains carried at least one whole Lpf operon (geneslpfABCDand/orlpfABCDE). The 64 enterohaemorrhagic (EHEC) and 24 enteropathogenic (EPEC) strains all carried two Lpf operons, allele 3 oflpfA1and allele 2 oflpfA2, a combination characteristic of the O157:H7/NM serotype. Out of the 9 bovine atypical (AT;stx-, eae-) strains, 7 carried one complete Lpf operon, allele 1 oflpfA2. The atypical strains belonged to main phylogenetic groups A and B1, while the EHEC and EPEC strains were from group D. Lpf variants carried by the 72 strains of theEscherichia coliReference Collection (ECOR) were determined with the same typing scheme. Alleles were detected in 25 strains, of which 6 were found negative for the respective Lpf operons in earlier studies. The marker value of the Lpf allelic combination for the O157:H7/NM serotype was confirmed, and further evidence was given for the presence of at least two different genetic lineages of atypical bovineE. coliO157 strains.

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Detection of virulence genes, phylogenetic group and antibiotic resistance of uropathogenic Escherichia coli in Mongolia

The Journal of Infection in Developing Countries ◽

10.3855/jidc.7903 ◽

2017 ◽

Vol 11 (01) ◽

pp. 51-57 ◽

Cited By ~ 17

Author(s):

Yandag Munkhdelger ◽

Nyamaa Gunregjav ◽

Altantsetseg Dorjpurev ◽

Nishi Juniichiro ◽

Jav Sarantuya

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Virulence Genes ◽

Wide Spectrum ◽

Iron Acquisition ◽

Uropathogenic Escherichia Coli ◽

Phylogenetic Group ◽

Diffusion Method ◽

Phylogenetic Groups ◽

E Coli

Introduction: The severity of urinary tract infection (UTI) produced by uropathogenic Escherichia coli (UPEC) is due to the expression of a wide spectrum of virulence genes. E. coli strains were divided into four phylogenetic groups (A, B1, B2 and D) based on their virulence genes. The present study aimed to assess the relationship between virulence genes, phylogenetic groups, and antibiotic resistance of UPEC. Methodology: A total of 148 E. coli were tested for antimicrobial resistance against 10 drugs using the disk diffusion method. The isolates were screened by polymerase chain reaction (PCR) for detection of virulence genes and categorized into the four major phylogenetic groups. Results: Phylogenetic group B2 was predominant (33.8%), followed by D (28.4%), A (19.6), and B1 (18.2%). A higher prevalence of fimH (89.9%), fyuA (70.3%), traT (66.2%), iutA (62.2%), kpsMTII (58.8%), and aer (56.1%) genes were found in UPEC, indicating a putative role of adhesins, iron acquisition systems, and protectins that are main cause of UTIs. The most common antibiotic resistance was to cephalotin (85.1%), ampicillin (78.4%) and the least to nitrofurantoin (5.4%) and imipenem (2%). In total, 93.9% of isolates were multidrug resistant (MDR). Conclusions: This study showed that group B2 and D were the predominant phylogenetic groups and virulence-associated genes were mostly distributed in these groups. The virulence genes encoding components of adhesins, iron acquisition systems, and protectins were highly prevalent among antibiotic-resistant UPEC. Although the majority of strains are MDR, nitrofurantoin is the drug of choice for treatment of UTI patients in Ulaanbaatar.

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Comparison of virulence factors and expression of specific genes between uropathogenic Escherichia coli and avian pathogenic E. coli in a murine urinary tract infection model and a chicken challenge model

Microbiology ◽

10.1099/mic.0.024869-0 ◽

2009 ◽

Vol 155 (5) ◽

pp. 1634-1644 ◽

Cited By ~ 58

Author(s):

Lixiang Zhao ◽

Song Gao ◽

Haixia Huan ◽

Xiaojing Xu ◽

Xiaoping Zhu ◽

...

Keyword(s):

Escherichia Coli ◽

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Virulence Genes ◽

Iron Acquisition ◽

Infection Model ◽

Phylogenetic Groups ◽

E Coli

Avian pathogenic Escherichia coli (APEC) and uropathogenic E. coli (UPEC) establish infections in extraintestinal habitats of different hosts. As the diversity, epidemiological sources and evolutionary origins of extraintestinal pathogenic E. coli (ExPEC) are so far only partially defined, in the present study,100 APEC isolates and 202 UPEC isolates were compared by their content of virulence genes and phylogenetic groups. The two groups showed substantial overlap in terms of their serogroups, phylogenetic groups and virulence genotypes, including their possession of certain genes associated with large transmissible plasmids of APEC. In a chicken challenge model, both UPEC U17 and APEC E058 had similar LD50, demonstrating that UPEC U17 had the potential to cause significant disease in poultry. To gain further information about the similarities between UPEC and APEC, the in vivo expression of 152 specific genes of UPEC U17 and APEC E058 in both a murine urinary tract infection (UTI) model and a chicken challenge model was compared with that of these strains grown statically to exponential phase in rich medium. It was found that in the same model (murine UTI or chicken challenge), various genes of UPEC U17 and APEC E058 showed a similar tendency of expression. Several iron-related genes were upregulated in the UTI model and/or chicken challenge model, indicating that iron acquisition is important for E. coli to survive in blood or the urinary tract. Based on these results, the potential for APEC to act as human UPEC or as a reservoir of virulence genes for UPEC should be considered. Further, this study compared the transcriptional profile of virulence genes among APEC and UPEC in vivo.

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Avian pathogenic Escherichia coli (APEC) and uropathogenic Escherichia coli (UPEC): characterization and comparison

The Journal of Infection in Developing Countries ◽

10.3855/jidc.14217 ◽

2021 ◽

Vol 15 (07) ◽

pp. 962-971

Author(s):

Daniela Tonini da Rocha ◽

Felipe De Oliveira Salle ◽

Karen Apellanis Borges ◽

Thales Quedi Furian ◽

Vladimir Pinheiro do Nascimento ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Susceptibility ◽

Antimicrobial Agents ◽

Resistance Rate ◽

Phylogenetic Groups ◽

Pathogenic Potential ◽

E Coli ◽

Antimicrobial Resistance Genes ◽

Tract Infections

Introduction: Avian pathogenic E. coli (APEC) and uropathogenic E. coli (UPEC) are responsible for avian colibacillosis and human urinary tract infections, respectively. There are genetic similarities between the APEC and UPEC pathotypes, suggesting the APEC strains could be a potential reservoir of virulence and antimicrobial-resistance genes for the UPEC strains. This study aimed to characterize and compare APEC and UPEC strains regarding the phylogroup classification, pathogenicity and antimicrobial susceptibility. Methodology: A total of 238 APEC and 184 UPEC strains were selected and characterized. The strains were assayed for antimicrobial susceptibility and classified into phylogenetic groups using a multiplex-PCR protocol. In addition, the APEC strains had previously been classified according to their in vivo pathogenicity. Results: The results showed that both pathotypes had variation in their susceptibility to most of the antimicrobial agents evaluated, with few strains classified as multidrug resistant. The highest resistance rate for both pathotypes was to amoxicillin. Classifying the APEC and UPEC strains into phylogenetic groups determined that the most frequently frequencies were for groups D and B2, respectively. These results reflect the pathogenic potential of these strains, as all the UPEC strains were isolated from unhealthy patients, and most of the APEC strains were previously classified as pathogenic. Conclusions: The results indicate that distribution into phylogenetic groups provided, in part, similar classification to those of in vivo pathogenicity index, as it was possible to adequately differentiate most of the pathogenic and commensal or low-pathogenicity bacteria. However, no relationship could be found between the specific antimicrobial agents and pathogenicity or phylogenetic group for either pathotype.

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Antimicrobial Resistance Genes and Diversity of Clones among ESBL- and Acquired AmpC-Producing Escherichia coli Isolated from Fecal Samples of Healthy and Sick Cats in Portugal

Antibiotics ◽

10.3390/antibiotics10030262 ◽

2021 ◽

Vol 10 (3) ◽

pp. 262

Author(s):

Isabel Carvalho ◽

Nadia Safia Chenouf ◽

Rita Cunha ◽

Carla Martins ◽

Paulo Pimenta ◽

...

Keyword(s):

Escherichia Coli ◽

Public Health Problem ◽

Phylogenetic Groups ◽

Disk Diffusion Test ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Specific Pcr ◽

Flight Mass Spectrometry ◽

Antimicrobial Resistance Genes ◽

Esbl Producers

The aim of the study was to analyze the mechanisms of resistance in extended-spectrum beta-lactamase (ESBL)- and acquired AmpC (qAmpC)-producing Escherichia coli isolates from healthy and sick cats in Portugal. A total of 141 rectal swabs recovered from 98 sick and 43 healthy cats were processed for cefotaxime-resistant (CTXR) E. coli recovery (in MacConkey agar supplemented with 2 µg/mL cefotaxime). The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) method was used for E. coli identification and antimicrobial susceptibility was performed by a disk diffusion test. The presence of resistance/virulence genes was tested by PCR sequencing. The phylogenetic typing and multilocus sequence typing (MLST) were determined by specific PCR sequencing. CTXRE. coli isolates were detected in seven sick and six healthy cats (7.1% and 13.9%, respectively). Based on the synergy tests, 11 of 13 CTXRE. coli isolates (one/sample) were ESBL-producers (ESBL total rate: 7.8%) carrying the following ESBL genes: blaCTX-M-1 (n = 3), blaCTX-M-15 (n = 3), blaCTX-M-55 (n = 2), blaCTX-M-27 (n = 2) and blaCTX-M-9 (n = 1). Six different sequence types were identified among ESBL-producers (sequence type/associated ESBLs): ST847/CTX-M-9, CTX-M-27, CTX-M-1; ST10/CTX-M-15, CTX-M-27; ST6448/CTX-M-15, CTX-M-55; ST429/CTX-M-15; ST101/CTX-M-1 and ST40/CTX-M-1. Three of the CTXR isolates were CMY-2-producers (qAmpC rate: 2.1%); two of them were ESBL-positive and one ESBL-negative. These isolates were typed as ST429 and ST6448 and were obtained in healthy or sick cats. The phylogenetic groups A/B1/D/clade 1 were detected among ESBL- and qAmpC-producing isolates. Cats are carriers of qAmpC (CMY-2)- and ESBL-producing E. coli isolates (mostly of variants of CTX-M group 1) of diverse clonal lineages, which might represent a public health problem due to the proximity of cats with humans regarding a One Health perspective.

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Iron Acquisition of Urinary Tract Infection Escherichia coli Involves Pathogenicity in Caenorhabditis elegans

Microorganisms ◽

10.3390/microorganisms9020310 ◽

2021 ◽

Vol 9 (2) ◽

pp. 310

Author(s):

Masayuki Hashimoto ◽

Yi-Fen Ma ◽

Sin-Tian Wang ◽

Chang-Shi Chen ◽

Ching-Hao Teng

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Caenorhabditis Elegans ◽

Large Scale ◽

Iron Acquisition ◽

Infection Model ◽

High Throughput Analysis ◽

E Coli ◽

C Elegans ◽

Tract Infections

Uropathogenic Escherichia coli (UPEC) is a major bacterial pathogen that causes urinary tract infections (UTIs). The mouse is an available UTI model for studying the pathogenicity; however, Caenorhabditis elegans represents as an alternative surrogate host with the capacity for high-throughput analysis. Then, we established a simple assay for a UPEC infection model with C. elegans for large-scale screening. A total of 133 clinically isolated E. coli strains, which included UTI-associated and fecal isolates, were applied to demonstrate the simple pathogenicity assay. From the screening, several virulence factors (VFs) involved with iron acquisition (chuA, fyuA, and irp2) were significantly associated with high pathogenicity. We then evaluated whether the VFs in UPEC were involved in the pathogenicity. Mutants of E. coli UTI89 with defective iron acquisition systems were applied to a solid killing assay with C. elegans. As a result, the survival rate of C. elegans fed with the mutants significantly increased compared to when fed with the parent strain. The results demonstrated, the simple assay with C. elegans was useful as a UPEC infectious model. To our knowledge, this is the first report of the involvement of iron acquisition in the pathogenicity of UPEC in a C. elegans model.

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The prevalence of the iutA and ibeA genes in Escherichia coli isolates from severe and non-severe patients with bacteremic acute biliary tract infection is significantly different

Gut Pathogens ◽

10.1186/s13099-021-00429-1 ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Mahoko Ikeda ◽

Tatsuya Kobayashi ◽

Fumie Fujimoto ◽

Yuta Okada ◽

Yoshimi Higurashi ◽

...

Keyword(s):

Escherichia Coli ◽

Tract Infection ◽

Biliary Tract ◽

Iron Acquisition ◽

Tertiary Care ◽

Care Hospital ◽

Biliary Tract Infection ◽

E Coli ◽

Severe Group ◽

Tract Infections

Abstract Background Although Escherichia coli is the most frequently isolated microorganism in acute biliary tract infections with bacteremia, data regarding its virulence are limited. Results Information on cases of bacteremia in acute biliary tract infection in a retrospective study was collected from 2013 to 2015 at a tertiary care hospital in Japan. Factors related to the severity of infection were investigated, including patient background, phylogenetic typing, and virulence factors of E. coli, such as adhesion, invasion, toxins, and iron acquisition. In total, 72 E. coli strains were identified in 71 cases, most of which primarily belonged to the B2 phylogroup (68.1%). The presence of the iutA gene (77.3% in the non-severe group, 46.4% in the severe group, P = 0.011) and the ibeA gene (9.1% in the non-severe group, and 35.7% in the severe group, P = 0.012) was significantly associated with the severity of infection. Among the patient characteristics, diabetes mellitus with organ involvement and alkaline phosphatase were different in the severe and non-severe groups. Conclusions We showed that bacteremic E. coli strains from acute biliary tract infections belonged to the virulent (B2) phylogroup. The prevalence of the iutA and ibeA genes between the two groups of bacteremia severity was significantly different.

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Phylogenetic Grouping and Virulence Potential of Extended-Spectrum-β-Lactamase-Producing Escherichia coli Strains in Cattle

Applied and Environmental Microbiology ◽

10.1128/aem.00351-12 ◽

2012 ◽

Vol 78 (13) ◽

pp. 4677-4682 ◽

Cited By ~ 38

Author(s):

Charlotte Valat ◽

Frédéric Auvray ◽

Karine Forest ◽

Véronique Métayer ◽

Emilie Gay ◽

...

Keyword(s):

Escherichia Coli ◽

Virulence Determinants ◽

Phylogenetic Groups ◽

Content Type ◽

E Coli ◽

Extended Spectrum ◽

Specific Distribution ◽

Extended Spectrum Beta Lactamases ◽

Phylogenetic Grouping ◽

Almost All

ABSTRACTIn line with recent reports of extended-spectrum beta-lactamases (ESBLs) inEscherichia coliisolates of highly virulent serotypes, such as O104:H4, we investigated the distribution of phylogroups (A, B1, B2, D) and virulence factor (VF)-encoding genes in 204 ESBL-producingE. coliisolates from diarrheic cattle. ESBL genes, VFs, and phylogroups were identified by PCR and a commercial DNA array (Alere, France). ESBL genes belonged mostly to the CTX-M-1 (65.7%) and CTX-M-9 (27.0%) groups, whereas those of the CTX-M-2 and TEM groups were much less represented (3.9% and 3.4%, respectively). One ESBL isolate wasstx1andeaepositive and belonged to a major enterohemorrhagicE. coli(EHEC) serotype (O111:H8). Two other isolates wereeaepositive butstxnegative; one of these had serotype O26:H11. ESBL isolates belonged mainly to phylogroup A (55.4%) and, to lesser extents, to phylogroups D (25.5%) and B1 (15.6%), whereas B2 strains were quasi-absent (1/204). The number of VFs was significantly higher in phylogroup B1 than in phylogroups A (P= 0.04) and D (P= 0.02). Almost all of the VFs detected were found in CTX-M-1 isolates, whereas only 64.3% and 33.3% of them were found in CTX-M-9 and CTX-M-2 isolates, respectively. These results indicated that the widespread dissemination of theblaCTX-Mgenes within theE. colipopulation from cattle still spared the subpopulation of EHEC/Shiga-toxigenicE. coli(STEC) isolates. In contrast to other reports on non-ESBL-producing isolates from domestic animals, B1 was not the main phylogroup identified. However, B1 was found to be the most virulent phylogroup, suggesting host-specific distribution of virulence determinants among phylogenetic groups.

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Antimicrobial susceptibility and pathogenicity of Escherichia coli strains of environmental origin

Ciência Rural ◽

10.1590/0103-8478cr20141020 ◽

2015 ◽

Vol 45 (7) ◽

pp. 1249-1255 ◽

Cited By ~ 5

Author(s):

Daiane Carvalho ◽

Fabrine Finkler ◽

Tiela Trapp Grassotti ◽

Hiran Castagnino Kunert Filho ◽

Francisco Esmaile de Sales Lima ◽

...

Keyword(s):

Escherichia Coli ◽

Antimicrobial Susceptibility ◽

Disc Diffusion ◽

Diffusion Test ◽

Pathogenic Potential ◽

Highly Pathogenic ◽

E Coli ◽

Intermediate Degree

The study aimed to evaluate the antimicrobial susceptibility of 109 samples of Escherichia coli (E. coli) of environmental origin and to characterize these isolates according to the degree of pathogenicity in vivo, verifying a possible relationship between this variable and susceptibility to the active principles tested. The isolates were subjected to disc diffusion test to 14 antibiotics. From 16.5% to 90% of the samples were sensitive; 1 - 28.5% showed intermediate degree of susceptibility and between 9 to 78% of E. coli analyzed were resistant. The highest resistance percentages were seen in the class of quinolones and tetracyclines (>75%), and for sensitivity in the class of amphenicols (68.8%). By inoculating 1- day - old chicks, the isolates were classified as highly pathogenic (2.7%), intermediate (10.1%), low (42.2%) and apathogenic (45%). It was observed a wide variation in the susceptibility profile of isolates in relation to antimicrobials. It was also found that most of the samples had pathogenic potential (55%), thus being considered as APEC (avian pathogenic E. coli). No relationship between pathogenicity and antimicrobial susceptibility (P≤0.05) was observed.

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Ferric Citrate Uptake is a Virulence Factor in Uropathogenic Escherichia coli

10.1101/2021.11.12.468464 ◽

2021 ◽

Author(s):

Arwen E Frick-Cheng ◽

Anna Sintsova ◽

Sara N Smith ◽

Ali Pirani ◽

Evan S Snitkin ◽

...

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Iron Acquisition ◽

Uropathogenic Escherichia Coli ◽

Ferric Citrate ◽

Uptake System ◽

Lipocalin 2 ◽

Compensatory Mechanisms ◽

E Coli ◽

Iron Source

More than half of women will experience a urinary tract infection (UTI) with uropathogenic Escherichia coli (UPEC) causing ~80% of uncomplicated cases. Iron acquisition systems are essential for uropathogenesis, and UPEC encode functionally redundant iron acquisition systems, underlining their importance. However, a recent UPEC clinical isolate, HM7 lacks this functional redundancy and instead encodes a sole siderophore, enterobactin. To determine if E. coli HM7 possesses unidentified iron acquisition systems, we performed RNA-sequencing under iron-limiting conditions and demonstrated that the ferric citrate uptake system (fecABCDE and fecIR) was highly upregulated. Importantly, there are high levels of citrate within urine, some of which is bound to iron, and the fec system is highly enriched in UPEC isolates compared to environmental or fecal strains. Therefore, we hypothesized that HM7 and other similar strains use the fec system to acquire iron in the host. Deletion of both enterobactin biosynthesis and ferric citrate uptake (ΔentB/ΔfecA) abrogates use of ferric citrate as an iron source and fecA provides an advantage in human urine in absence of enterobactin. However, in a UTI mouse model, fecA is a fitness factor independent of enterobactin production, likely due to the action of host Lipocalin-2 chelating ferrienterobactin. These findings indicate that ferric citrate uptake is used as an iron source when siderophore efficacy is limited, such as in the host during UTI. Defining these novel compensatory mechanisms and understanding the nutritional hierarchy of preferred iron sources within the urinary tract are important in the search for new approaches to combat UTI.

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