The Potential Virulence Factors of<i>Providencia stuartii</i>: Motility, Adherence, and Invasion

Kurmasheva, Naziia; Vorobiev, Vyacheslav; Шарипова, М. Р.; Ефремова, Т. В.; Марданова, А. М.

doi:10.1155/2018/3589135

Cited by 32 publications

(21 citation statements)

References 45 publications

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“…Despite the clinical significance of this organism, no previous studies explored P. stuartii genes essential for growth in vitro and little is known regarding P. stuartii virulence factors. Adherence, biofilm formation, host cell invasion, and motility are important aspects of urinary tract colonization for numerous uropathogenic bacteria, and in vitro and in silico evidence suggests that these properties may contribute to P. stuartii pathogenesis (20)(21)(22)(23)(24)(25)(26). However, their contribution to colonization and pathogenesis has not been assessed experimentally.…”

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confidence: 99%

Transposon Insertion Site Sequencing of Providencia stuartii: Essential Genes, Fitness Factors for Catheter-Associated Urinary Tract Infection, and the Impact of Polymicrobial Infection on Fitness Requirements

Johnson

Forsyth

Smith

et al. 2020

mSphere

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Providencia stuartii is a common cause of polymicrobial catheter-associated urinary tract infection (CAUTI), and yet literature describing the molecular mechanisms of its pathogenesis is limited. To identify factors important for colonization during single-species infection and during polymicrobial infection with a common cocolonizer, Proteus mirabilis, we created a saturating library of ∼50,000 transposon mutants and conducted transposon insertion site sequencing (Tn-Seq) in a murine model of CAUTI. P. stuartii strain BE2467 carries 4,398 genes, 521 of which were identified as essential for growth in laboratory medium and therefore could not be assessed for contribution to infection. Using an input/output fold change cutoff value of 20 and P values of <0.05, 340 genes were identified as important for establishing single-species infection only and 63 genes as uniquely important for polymicrobial infection with P. mirabilis, and 168 genes contributed to both single-species and coinfection. Seven mutants were constructed for experimental validation of the primary screen that corresponded to flagella (fliC mutant), twin arginine translocation (tatC), an ATP-dependent protease (clpP), d-alanine-d-alanine ligase (ddlA), type 3 secretion (yscI and sopB), and type VI secretion (impJ). Infection-specific phenotypes validated 6/7 (86%) mutants during direct cochallenge with wild-type P. stuartii and 3/5 (60%) mutants during coinfection with P. mirabilis, for a combined validation rate of 9/12 (75%). Tn-Seq therefore successfully identified genes that contribute to fitness of P. stuartii within the urinary tract, determined the impact of coinfection on fitness requirements, and added to the identification of a collection of genes that may contribute to fitness of multiple urinary tract pathogens. IMPORTANCE Providencia stuartii is a common cause of polymicrobial catheter-associated urinary tract infections (CAUTIs), particularly during long-term catheterization. However, little is known regarding the pathogenesis of this organism. Using transposon insertion site sequencing (Tn-Seq), we performed a global assessment of P. stuartii fitness factors for CAUTI while simultaneously determining how coinfection with another pathogen alters fitness requirements. This approach provides four important contributions to the field: (i) the first global estimation of P. stuartii genes essential for growth in laboratory medium, (ii) identification of novel fitness factors for P. stuartii colonization of the catheterized urinary tract, (iii) identification of core fitness factors for both single-species and polymicrobial CAUTI, and (iv) assessment of conservation of fitness factors between common uropathogens. Genomewide assessment of the fitness requirements for common uropathogens during single-species and polymicrobial CAUTI thus elucidates complex interactions that contribute to disease severity and will uncover conserved targets for therapeutic intervention.

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confidence: 99%

Transposon Insertion Site Sequencing of Providencia stuartii: Essential Genes, Fitness Factors for Catheter-Associated Urinary Tract Infection, and the Impact of Polymicrobial Infection on Fitness Requirements

Johnson

Forsyth

Smith

et al. 2020

mSphere

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“…Pretreatment of DA with DTT increases the detected relative abundance of many Enterobacteriaceae members, including the clinically important genera Klebsiella, Providencia and Salmonella as well as unknown members. Providencia and Salmonella include motile species that can adhere to mucus and epithelial cells and actively invade the host epithelium [27][28][29]. The relative abundance of the genus Pseudomonas, detected in DA-DTT was also increased compared to non-pretreated DA.…”

Section: Discussionmentioning

confidence: 99%

Optimizing microbiome sequencing for small intestinal aspirates: validation of novel techniques through the REIMAGINE study

et al. 2019

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Background: The human small intestine plays a central role in the processes of digestion and nutrient absorption. However, characterizations of the human gut microbiome have largely relied on stool samples, and the associated methodologies are ill-suited for the viscosity and low microbial biomass of small intestine samples. As part of the REIMAGINE study to examine the specific roles of the small bowel microbiome in human health and disease, this study aimed to develop and validate methodologies to optimize microbial analysis of the small intestine. Results: Subjects undergoing esophagogastroduodenoscopy without colon preparation for standard of care were prospectively recruited, and~2 ml samples of luminal fluid were obtained from the duodenum using a custom sterile aspiration catheter. Samples of duodenal aspirates were either untreated (DA-U, N = 127) or pretreated with dithiothreitol (DA-DTT, N = 101), then cultured on MacConkey agar for quantitation of aerobic gram-negative bacteria, typically from the class Gammaproteobacteria, and on blood agar for quantitation of anaerobic microorganisms. DA-DTT exhibited 2.86-fold greater anaerobic bacterial counts compared to DA-U (P = 0.0101), but were not statistically different on MacConkey agar. DNA isolation from DA-U (N = 112) and DA-DTT (N = 43) samples and library preparation for 16S rRNA gene sequencing were also performed using modified protocols. DA-DTT samples exhibited 3.81-fold higher DNA concentrations (P = 0.0014) and 4.18-fold higher 16S library concentrations (P < 0.0001) then DA-U samples. 16S rRNA gene sequencing revealed increases in the detected relative abundances of obligate and facultative anaerobes in DA-DTT samples, including increases in the genera Clostridium (false discovery rate (FDR) P = 4.38E-6), Enterococcus (FDR P = 2.57E-8), Fusobacterium (FDR P = 0.02) and Bacteroides (FDR P = 5.43E-9). Detected levels of Gram-negative enteropathogens from the phylum Proteobacteria, such as Klebsiella (FDR P = 2.73E-6) and Providencia (FDR P < 0.0001) (family Enterobacteriaceae) and Pseudomonas (family Pseudomonadaceae) (FDR P = 0.04), were also increased in DA-DTT samples. Conclusions: This study validates novel DTT-based methodology which optimizes microbial culture and 16S rRNA gene sequencing for the study of the small bowel microbiome. The microbial analyses indicate increased isolation of facultative and obligate anaerobes from the mucus layer using these novel techniques.

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“…Providencia stuartii is a Gram negative bacterium which is usually found in soil, sewage and water. It is a member of Enterobacteriaceae family and known to cause nosocomial infections [14]. Pantoea dispersa is a member of Enterobacteriaceae family that is found in soil, water and plants and seldom causes human infections but was in recent times associated with neonatal sepsis as a causal agent [15].…”

Section: Discussionmentioning

confidence: 99%

Effect of Heavy Metals on β-galactosidase Activity in Marine Bacteria

Osigwe

Ariole

Ibiene

2020

JAMB

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Background: Due to metal pollution and its toxicity in the coastal areas, the enzymatic activities of bacteria involved in the breakdown of organic compounds are repressed leading to decline in biodegradation rate. Aim: The influence of heavy metals (copper, lead, zinc, manganese and iron) on β-galactosidase activity in three bacterial strains (Providencia stuartii, Pantoea dispersa and Aeromonas dhakensis) isolated from coastal marine sediment collected from coastal zone in Bonny Island, Nigeria was investigated. Methodology: The strains were cultivated in Z- buffered medium having lactose as enzyme inducer. Beta galactosidase assay was done via 2-nitrophenol β-D-galactopyranoside as the substrate. The absorbances of p-nitrophenol solution formed were measured at 420 nm in a spectrophotometer. The β-galactosidase activities were calculated comparative to controls. Results: Presence of the metals significantly affected β-galactosidase activities. Metal concentration of 0.001 mg/L triggered a decrease in enzyme activity. The sensitivity patterns of Pantoea dispersa and Aeromonas dhakensis were Cu>Pb>Zn>Mn>Fe while that of Providencia stuartii was Pb>Cu>Zn>Mn>Fe. The effect of metal stress to enzyme synthesis is reliant on the organism and the metal. This might be described with logistic dose-response model using elevated coefficient of inhibition (R > 0.81). Conclusion: The results revealed that concentration of metal as low as 0.001 mg/L when deposited in the environment has detrimental effect on microbial activities and consequently on biogeochemical cycles. The isolated bacterial strains could serve as ideal organisms for heavy metal toxicity evaluation.

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The Potential Virulence Factors ofProvidencia stuartii: Motility, Adherence, and Invasion

Cited by 32 publications

References 45 publications

Transposon Insertion Site Sequencing of Providencia stuartii: Essential Genes, Fitness Factors for Catheter-Associated Urinary Tract Infection, and the Impact of Polymicrobial Infection on Fitness Requirements

Transposon Insertion Site Sequencing of Providencia stuartii: Essential Genes, Fitness Factors for Catheter-Associated Urinary Tract Infection, and the Impact of Polymicrobial Infection on Fitness Requirements

Optimizing microbiome sequencing for small intestinal aspirates: validation of novel techniques through the REIMAGINE study

Effect of Heavy Metals on β-galactosidase Activity in Marine Bacteria

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