Human Urinary Tract Infections Research Articles

Comparative in vitro Study of Antimicrobial, Antibiofilm and Quorum Sensing Inhibitory Activities of Hypericum calycinum L. and Parietaria officinalis L. Extracts

Aim: This study aims to elucidate both the anti-virulence and antimicrobial effects of ethanol extracts from Hypericum calycinum L. and Parietaria officinalis L. Material and Methods: Antimicrobial activity was evaluated using the well diffusion method against five bacteria and two yeast isolates involved in human urinary tract infections (UTIs). The potential of the extracts to inhibit quorum sensing (QS), was assessed using the biosensor strain Chromobacterium violaceum ATCC 12472. Additionally, the antibiofilm activities were investigated using a microplate biofilm assay on Escherichia coli and Pseudomonas aeruginosa. Results: H. calycinum exhibited the highest inhibitory effect at a concentration of 100 mg/mL against Candida albicans with an inhibition zone of 24.5±0.71 mm, while P. officinalis showed its highest effect at the same concentration against E. coli with an inhibition zone of 15.5±0.71 mm. Overall, H. calycinum demonstrated stronger antimicrobial activity compared to P. officinalis. Both plant extracts inhibited QS at similar levels, with inhibition zones ranging between 10-12 mm. The antibiofilm effect varied depending on the bacterial species, but notably, P. officinalis extract exhibited over 80% antibiofilm efficacy against E. coli at all concentrations. Conclusion: This study demonstrates that H. calycinum L. and P. officinalis L. are potent antimicrobial agents against UTI pathogens. While their anti-QS efficacy is not exceptional, the significant inhibition of E. coli biofilm formation underscores their potential as formidable agents. Designed as a fundamental study, it highlights the promising antimicrobial properties of these plant extracts and marks the first investigation into their capabilities as QS and biofilm-preventive agents.

The infectious capacity of Enterococcus faecalis, Staphylococcus aureus, and Staphylococcus saprophyticus in a porcine model of urinary tract infection.

The purpose of this study was to establish a porcine model of urinary tract infection (UTI) with gram-positive uropathogens. Ten female domestic pigs were experimentally inoculated with human UTI isolates of Enterococcus faecalis (n = 3), Staphylococcus saprophyticus (n = 3), or Staphylococcus aureus (n = 4) and followed with regular urine samples. Bladders and kidneys were aseptically removed at termination (5-7 days post infection) and assessed by gross pathology and bacterial enumeration. Enterococcus faecalis (n = 3 of 3) and S. aureus (n = 2 of 4) successfully colonized the pig bladders. Inoculation with S. saprophyticus never resulted in detectable bacteriuria. All infected pigs had cleared the infection spontaneously before termination. Surprisingly, three (of four) pigs inoculated with S. aureus led to spontaneous infection with opportunistic pathogens. Also, one pig colonized with E. faecalis resulted in spontaneous infection with E. coli. In conlusion, the pig supports experimental UTI with E. faecalis for up to 24 h but not prolonged infection. S. aureus and S. saprophyticus fails to cause UTI in pigs and other animals should be considered for studying these pathogens.

Genomic Analysis of Enterobacter Species Isolated from Patients in United States Hospitals.

We analyzed the whole genome sequences (WGS) and antibiograms of 35 Enterobacter isolates, including E. hormaechei and E. asburiae, and the recently described E. bugandensis, E. kobei, E. ludwigii, and E. roggenkampii species. Isolates were obtained from human blood and urinary tract infections in patients in the United States. Our goal was to understand the genetic diversity of antimicrobial resistance genes and virulence factors among the various species. Thirty-four of 35 isolates contained an AmpC class blaACT allele; however, the E. roggenkampii isolate contained blaMIR-5. Of the six Enterobacter isolates resistant to ertapenem, imipenem, and meropenem, four harbored a carbapenemase gene, including blaKPC or blaNDM. All four isolates were mCIM-positive. The remaining two isolates had alterations in ompC genes that may have contributed to the resistance phenotype. Interpretations of cefepime test results were variable when disk diffusion and automated broth microdilution results were compared due to the Clinical Laboratory and Standards Institute use of the "susceptible dose-dependent" classification. The diversity of the blaACT alleles paralleled species identifications, as did the presence of various virulence genes. The classification of recently described Enterobacter species is consistent with their resistance gene and virulence gene profiles.

Diversity and Antibiotic Susceptibility of Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae from the Urine of Human Population Using Bacteria-Contaminated Water in Butembo Urban Area (DR Congo, Central Africa)

Aims: This study aims to investigate the extended-spectrum beta-lactamases (ESBLs)-producing Enterobacteriaceae in the human urine samples and to understand the potential influence of the contaminated water the urine donors use. Study Design and Methodology: One hundred urine samples from females and the same number from males were collected from donors of different age groups and analysed. One hundred urine samples from females and the same number from males were collected from donors of different age groups. The enterobacterial isolation was done on CLED agar medium. Their identification was carried out first by the enzymatic technic, then using MALDI–TOF MS method. ESBLs-producing enterobacteria were identified by searching the synergy between Clavulanic acid and 3rd generation Cephalosporins. The antimicrobial susceptibility tests were carried out using the disk diffusion method. Results: In male urine samples, the identified ESBLs-producing strains belong to E. coli, E. agglomerans and K. pneumoniae. They were in 11 samples: 3 from donors aged 20 years or less, 3 also from those aged 21 to 40 years as well as in those aged 41 to 60 years, and 2 in samples from donors aged over than 60 years. In female urine samples, the identified ESBLs-producing strains belong to Escherichia coli, Enterobacter agglomerans, Yersinia frederiksenii, Salmonella typhi and Klebsiella pneumoniae. They were in 27 samples: 3 from donors aged 20 years or less, 9 from those aged 21 to 40 years, 12 from those aged 41 to 60 years, and 3 from donors aged over than 60 years. All the strains were sensitive to Imipenem and resistant to Cefotaxim, Ceftazidin, Amoxicillin+Clavulanic acid and Nitrofurantoin. The Multi-Antibiotic Resistance (MAR) index varied from 0.416 (E. coli strains, youngest male donors) to 0.916 (E. coli strains, old female donors). Most of strains react differently from one antibiotic to another (P<0.05), with the exception of E. coli strains from most samples. Most of the identified ESBLs-producing species have been reported in surface and groundwater the population use. These water resources could play a role in the human urinary tract infections. Conclusion: There is a serious health problem with great MAR indices for all bacterial species identified. Treating water before use could reduce the viability and spreading of ESBLs-producing bacteria and genes into the population.

Association between antimicrobial usage in livestock and antimicrobial resistance in Escherichia coli isolates from human urinary tract infections in the Netherlands, 2009-2020.

In the last decade, veterinary antimicrobial usage (AMU) and antimicrobial resistance (AMR) among indicator bacteria in livestock have decreased substantially in the Netherlands. The extent to which this decrease has affected AMR levels among human infections remains unclear. To assess the association between AMU in livestock and AMR in Escherichia coli isolates from human urinary tract infections (UTIs). Data on AMR and AMU between 2009 and 2020 from Dutch national surveillance programmes for humans and livestock were used. Associations between AMU in four major livestock sectors and AMR in humans were assessed for 10 antimicrobial classes and the ESBL resistance profile, using logistic regression analysis. Associations between AMU and AMR in livestock, between AMR in livestock and in humans, and between AMU and AMR in humans were also assessed. Statistical significance was reached for 16/31 of the tested associations between AMU in livestock and AMR in human E. coli UTIs. Of the significant associations, 11 were positive (OR 1.01-1.24), whereas 5 were negative (OR 0.96-0.99). All associations between human AMU and AMR in E. coli isolates from UTIs were positive and statistically significant. Weak but significant positive correlations were also observed between livestock AMR and human AMR. Although several significant associations between AMU in livestock and AMR in human UTIs caused by E. coli were observed, the associations between AMU and AMR were generally stronger within the human and animal populations. This indicates that potential zoonotic spread of AMR in E. coli causing human UTIs from livestock sources is limited.

Colistin-resistance genes in Escherichia coli isolated from patients with urinary tract infections.

The incidence of antimicrobial resistance is alarmingly high because it occurs in humans, environment, and animal sectors from a "One Health" viewpoint. The emergence of plasmid-carried mobile colistin-resistance (MCR) genes limits the efficacy of colistin, which is the last-line treatment for multidrug resistance (MDR) against gram-negative infections. The current study aimed to investigate emergence of colistin-resistance (MCR 1-5) genes in E. coli isolated from patients with urinary tract infections (UTIs) in Jordan. E. coli (n = 132) were collected from urine specimens. The E. coli isolated from human UTI patients were examined the resistance to colistin based on the presence of MCR (1-5). All isolates were tested against 20 antimicrobials using the standard disk diffusion method. The broth microdilution technique was used to analyze colistin resistance. In addition, the MCR (1-5) genes were detected using multiplex PCR. Out of the 132 isolates, 1 isolate was colistin-resistant, having a minimum inhibitory concentration of 8 μg/mL and possessing MCR-1. All the E. coli isolates showed high resistance to penicillin (100%), amoxicillin (79.55%), cephalexin (75.76%), nalidixic acid (62.88%), tetracycline (58.33%), or cefepime (53.79). To our knowledge, this is the first report on the presence of plasmid-coded MCR-1 in E. coli from a patient with UTIs in Jordan. This is a problematic finding because colistin is the last-line drug for the treatment of infections caused by MDR gram-negative bacteria. There is a crucial need to robustly utilize antibiotics to control and prevent the emergence and prevalence of colistin-resistance genes.

Prevalence and Antimicrobial Resistance Pattern of Uropathogenic Escherichia coli at a Tertiary Care Hospital: A Retrospective Study

Background: Uropathogenic Escherichia coli (UPEC) is the most common cause of urinary tract infection in humans including cystitis and pyelonephritis. Antimicrobial resistance (AMR) in UPEC is one of the global public challenges that is due to prevalent use of antibiotics in healthcare setting. Therefore, the purpose of this study is to investigate the frequency and antibiotic resistance pattern of UPEC isolated from patients admitted to Modares Hospital in Saveh, Iran. Methods: In this study, in total 633 isolates were evaluated. UPEC isolates were obtained from patients with urinary tract infection and identified using conventional microbiological protocols. Antibiotic resistance pattern of UPEC against different antibiotic were determined using disk diffusion method. SPSSTM software was used for statistical analysis Results: In this study, the most sample was related to outpatients and the lowest sample was related to the CCU wards. The highest antibiotic resistance showed against cephalothin (63.8%) and nalidixic acid (62.2%) antibiotics. The highest effective antibiotics for the tested UPEC was nitrofurantoin (90.7%) and gentamicin (77.3%). Cephalothin and nalidixic acid in hospitalized patients in ICU and emergency wards, respectively, showed the highest antibiotic resistance. Out of 633 UPEC, the rate of Multi Drug Resistant (MDR) isolates were 343 (54.2%). Conclusion: The result of this study highlighted the role of UPEC as one of the important cause of UTI in individuals. Also, nitrofurantoin then gentamicin are the most effective antibiotics against UPEC infections. Logical prescription of antibiotics and infection control strategies are needed for prevention and control of nosocomial infections especially urinary tract infection.

Bacterial diversity associated with urinary tract infections in humans

Bacterial diversity associated with urinary tract infections in humans

Draft genomic sequence for uropathogenic Staphylococcus saprophyticus ATCC 49453.

Staphylococcus saprophyticus, a common contaminant of foods, causes urinary tract infections in humans. Here, we report the draft genomic sequence for S. saprophyticus ATCC 49453, which is currently being used in food safety research.

Combined functional genomic and metabolomic approaches identify new genes required for growth in human urine by multidrug-resistant Escherichia coli ST131.

Uropathogenic Escherichia coli (UPEC) cause ~80% of all urinary tract infections (UTIs), with increasing rates of antibiotic resistance presenting an urgent threat to effective treatment. To cause infection, UPEC must grow efficiently in human urine (HU), necessitating a need to understand mechanisms that promote its adaptation and survival in this nutrient-limited environment. Here, we used a combination of functional genomic and metabolomic techniques and identified roles for the metabolism of small peptides, amino acids, nucleotides, and L-lactate, as well as the stringent response pathway, lipopolysaccharide biosynthesis, and fluoride resistance, for UPEC growth in HU. We further demonstrated that pathways involving nucleotide metabolism and the stringent response are required for UPEC colonization of the mouse bladder. The UPEC genes and metabolic pathways identified in this study represent targets for the development of innovative therapeutics to prevent UPEC growth during human UTI, an urgent need given the rapidly rising rates of global antibiotic resistance.

Key biological processes and essential genes for Proteus mirabilis biofilm development inhibition by protocatechuic acid

Proteus mirabilis is an opportunistic pathogen linked to human urinary tract infections, and is potentially present as a foodborne pathogen within poultry products, including broiler chickens. This report outlines the inhibitory impacts of protocatechuic acid (PCA) on P. mirabilis isolated from a broiler slaughterhouse in China as well as its biofilm. This investigation encompasses assays related to motility and adhesion, bacterial metabolic activity, extracellular polymer (EPS) production, and scavenging capacity. The findings demonstrated that PCA reduced biofilm formation by 61 %. Transcriptomics findings identified that PCA limited the expression of genes like PstS that promote adhesin formation, rbsA and RcsB that alter bacterial chemotaxis, lipopolysaccharide synthesis genes LpxA and EptB, and cell wall synthesis genes MurF and MrdA, and affects the Regulator of Capsule Synthesis (RCS) two-component modulation system. Weighted gene co-expression network analysis (WGCNA) was conducted to identify the core genes. Furthermore, the binding sites of PCA to cytochrome oxidases cydA and cydB, two subunits of ATP synthase atpI and atpH, and ftsZ, which regulate bacterial division, were predicted via molecular docking. Metabolome analysis determined that PCA critically influenced coenzyme A biosynthesis, nucleotide metabolism, alanine, aspartic acid, and glutamate metabolic pathways of P. mirabilis. Therefore, PCA impacts metabolism within bacteria via various pathways, limiting the levels of extracellular polymer and bacterial viability to hinder biofilm formation. Additionally, we prepared an antibacterial plastic film containing protocatechuic acid using PVA as the monomer and CNC as the reinforcing agent. We examined the mechanical and antibacterial properties of this film. When used to wrap chicken, it reduced the total number of colonies, slowed the deterioration of chicken, and maintained the freshness of chicken. In conclusion, the information outlined in this study complements our comprehension of P. mirabilis inhibition by PCA and provides clues for the reduction of foodborne infections associated with P. mirabilis.

Biofilm-forming ability, antibiotic resistance and phylogeny of Escherichia coli isolated from extra intestinal infections of humans, dogs, and chickens

Escherichia coli (E. coli) causes various infections in humans and animals. The biofilm-forming ability of E. coli has increased antimicrobial resistance and capacity to cause recurrent and chronic infections. This study determined the biofilm-forming ability of E. coli isolated from extraintestinal infections of humans, chickens, and dogs in relation to the phylogroup, type of infection, and antibiotic resistance. Isolates from chickens showed significantly higher biofilm-forming ability compared to those causing urinary tract infections in humans (p = 0.0001). Further, isolates belonging to phylogroup B1 displayed a higher likelihood to form biofilms. Resistance to ciprofloxacin and trimethoprim-sulfamethoxazole was positively correlated with biofilm-forming ability. Harbouring plasmid-mediated quinolone resistance gene, qnrS was also positively correlated with biofilm formation. This study provides insight into factors such as phylogroup and the type of infections that could enhance biofilm formation, as well as genotypic and phenotypic antibiotic resistance that could correlate with the ability to form biofilms.

In-Silico Screening for Vaccine Candidate Peptides within UPEC-UTI89 Strain against Urinary Tract Infection in Humans

In-Silico Screening for Vaccine Candidate Peptides within UPEC-UTI89 Strain against Urinary Tract Infection in Humans

Molecular study of hpmA and ureR genes among Proteus mirabilis isolates inpatient with urinary tract infection

The most frequently occurring human bacterial infections are urinary tract infections (UTIs). P. mirabilis is one of the common causes of UTIs in humans with complicated UTIs, the formation of stones in the kidney, and long-term indwelling catheters. This study aims to isolate, and identify Proteusmirabilis from patients with UTIs using traditional methods and 16s rRNA gene sequencing techniques, as well as detect some virulence genes (hpm A and ureR) in Proteus mirabilis by using polymerase chain reaction (PCR). A total of five isolates of Proteus mirabilis were isolated from patients with urinary tract infections in June 2022. Identification of isolates depends on microscopic and cultural characteristics and traditional biochemical tests, finally, isolates are identified by target gene 16s rRNA using PCR technique, in addition, this study investigates the genes responsible for virulence factors such as hpmA and hpmR.In this study laboratory diagnosis and molecular analysis (16s rRNA gene) revealed that the five isolates are Proteus mirabilis also this study shows the presence of the hpmAgenes by PCR techniques in the isolates 100%. Our study has shown that 16s rRNA was a strong eclectic power and an excellent method for the identification of bacteria also these study frequency ofureR and hpmA in P.mirabilis isolated from UTIs and these genes have a manifest role in the infection.

Detection of Extended-Spectrum β-Lactamase (ESBL) E. coli at Different Processing Stages in Three Broiler Abattoirs.

The European Food Safety Authority (EFSA) identified extended-spectrum β-lactamase/AmpC β-lactamase (ESBL/AmpC)-producing E. coli as one of the main priority hazards for poultry. Different studies detected ESBL-producing E. coli at broiler fattening farms and in abattoirs, concluding that poultry meat is a potential source of human infection. Broiler breast skin samples taken in three abattoirs with different scalding techniques were examined for ESBL-producing Escherichia (E.) coli and their phylogenetic groups. A total of 307 ESBL-producing E. coli isolates were found, and the abattoir with conventional immersion scalding with thermal treatment of the water had the lowest incidence. Phylogroups D/E and B1 were mostly detected, while phylogroups C, D, and E were not detected. Phylogroup B2 was detected in low proportions. The phylogroups B2 and D are important as they have been associated with urinary tract infections in humans, but were only detected in low proportions at different processing stages in this study. Since the risk for the consumer of being infected via chicken meat with ESBL-producing E. coli and E. coli of highly pathogenic phylogroups cannot be excluded, good kitchen hygiene is of great importance.

Draft genome sequences of two Proteus mirabilis isolates recovered from a municipal wastewater treatment plant in Ontario, Canada.

Proteus mirabilis is a Gram-negative bacterium that is frequently implicated in urinary tract infections in humans and companion animals and has also been associated with foodborne infections in several countries. Here, we report the draft genome sequences of two P. mirabilis isolates recovered from municipal wastewater.

Dogs Can Be Reservoirs of Escherichia coli Strains Causing Urinary Tract Infection in Human Household Contacts.

This study aimed to investigate the role played by pets as reservoirs of Escherichia coli strains causing human urinary tract infections (UTIs) in household contacts. Among 119 patients with community-acquired E. coli UTIs, we recruited 19 patients who lived with a dog or a cat. Fecal swabs from the household pet(s) were screened by antimicrobial selective culture to detect E. coli displaying the resistance profile of the human strain causing UTI. Two dogs shed E. coli isolates indistinguishable from the UTI strain by pulsed-field gel electrophoresis. Ten months later, new feces from these dogs and their owners were screened selectively and quantitatively for the presence of the UTI strain, followed by core-genome phylogenetic analysis of all isolates. In one pair, the resistance phenotype of the UTI strain occurred more frequently in human (108 CFU/g) than in canine feces (104 CFU/g), and human fecal isolates were more similar (2-7 SNPs) to the UTI strain than canine isolates (83-86 SNPs). In the other pair, isolates genetically related to the UTI strain (23-40 SNPs) were only detected in canine feces (105 CFU/g). These results show that dogs can be long-term carriers of E. coli strains causing UTIs in human household contacts.

Modeling the Temperature Effect on the Growth of Uropathogenic Escherichia coli in Sous-Vide Chicken Breast.

Uropathogenic Escherichia coli (UPEC) is known to cause 65-75% of human urinary tract infection (UTI) cases. Poultry meat is a reservoir of UPEC, which is suspected to cause foodborne UTIs. In the present study, we aimed to determine the growth potential of UPEC in ready-to-eat chicken breasts prepared by sous-vide processing. Four reference strains isolated from the urine of UTI patients (Bioresource Collection and Research Center [BCRC] 10,675, 15,480, 15,483, and 17,383) were tested by polymerase chain reaction assay for related genes to identify their phylogenetic type and UPEC specificity. A cocktail of these UPEC strains was inoculated into sous-vide cooked chicken breast at 103-4 colony-forming unit (CFU)/g and stored at 4°C, 10°C, 15°C, 20°C, 30°C, and 40°C. Changes in the populations of UPEC during storage were analyzed by a one-step kinetic analysis method using the U.S. Department of Agriculture [USDA] Integrated Pathogen Modeling Program-Global Fit [IPMP-Global Fit]. The results showed that the combination of the no lag phase primary model and the Huang square-root secondary model fitted well with the growth curves to obtain the appropriate kinetic parameters. This combination for predicting UPEC growth kinetics was further validated using it to study additional growth curves at 25°C and 37°C, which showed that the root mean square error, bias factor, and accuracy factor were 0.49-0.59 (log CFU/g), 0.941-0.984, and 1.056-1.063, respectively. In conclusion, the models developed in this study are acceptable and can be used to predict the growth of UPEC in sous-vide chicken breast.

Benzoic acid derivatives as potent antibiofilm agents against Klebsiella pneumoniae biofilm

Klebsiella pneumoniae is responsible for a significant proportion of human urinary tract infections, and its biofilm is a major virulence. One potential approach to controlling biofilm-associated infections is targeting the adhesin MrkD1P to disrupt biofilm formation. We employed Schrodinger's Maestro tool with the OPLS 2005 force field to dock compounds with the target protein. Two benzoic acid derivatives, 3-hydroxy benzoic acid and 2,5-dihydroxybenzoic acid, had strong binding free energies (-55.57 and-18.68kcal/mol) and were the most potent compounds. The in-vitro experiments were conducted to validate the in-silico results. The results showed that both compounds effectively inhibited biofilm formation at low concentrations (4 and 8mg/mL, respectively) and had antibiofilm activity, restricting cell attachment. Both compounds demonstrated a strong biofilm inhibitory effect, with 97% and 89% reduction in biofilm by 3-hydroxy benzoic acid and 2,5-dihydroxybenzoic acid, respectively. These findings suggest that natural compounds can be a potential source of new drugs to combat biofilm-associated infections. The study highlights the potential of targeting adhesin MrkD1P as an effective approach to controlling biofilm-associated infections caused by K.pneumoniae. The results may have implications for the development of new therapies for biofilm-associated infections and pave the way for future research in this area.

Assessing the urinary concentration of nitrofurantoin and its antibacterial activity against Escherichia coli, Staphylococcus pseudintermedius, and Enterococcus faecium isolated from dogs with urinary tract infections.

Nitrofurantoin, a broad-spectrum nitrofuran class antibiotic, is applied as a first-line antibiotic in treating human urinary tract infections (UTIs) due to its great efficacy and high achievable concentration. The interest in using this antibiotic in companion animals has increased due to the growing demand for effective antibiotics to treat UTIs caused by multidrug-resistant bacteria. Currently, the susceptibility interpretations for nitrofurantoin are based on the breakpoints set for humans, while the canine-specific breakpoints are still unavailable. In this study, we assessed the concentration of nitrofurantoin reaching the dog's urine using the recommended oral dosing regimen. In addition, we examined the efficacy of this breakpoint concentration against the common canine UTI pathogens, Escherichia coli, Staphylococcus pseudintermedius, and Enterococcus faecium. Eight experimental beagle dogs were treated with ~5 mg/kg of nitrofurantoin macrocrystal PO 8qh for 7 days. The urine samples were collected via cystocentesis at 2, 4, and 6 h after administration on day 2 and day 7 and used to quantify nitrofurantoin concentrations by ultra-high performance liquid chromatography. The results showed that 26.13-315.87 μg/mL nitrofurantoin was detected in the dogs' urine with a mean and median concentration of 104.82 and 92.75 μg/mL, respectively. Additionally, individual dogs presented with urinary nitrofurantoin concentrations greater than 64 μg/mL for at least 50% of the dosing intervals. This concentration efficiently killed E. coli, and S. pseudintermedius, but not E. faecium strains carrying an MIC90 value equal to 16, 16, and 128 μg/mL, respectively. Taken together, these results suggest that the value of 64 μg/mL may be set as a breakpoint against UTI pathogens, and nitrofurantoin could be an effective therapeutic drug against E. coli and S. pseudintermedius for canine UTIs.