Synovial Fluid Mediated Aggregation of Clinical Strains of Four Enterobacterial Species

Macías-Valcayo, Alicia; Staats, Amelia; Aguilera-Correa, John Jairo; Brooks, Jack; Gupta, Tripti Thapa; Dusane, Devendra H.; Stoodley, Paul; Esteban, Jaime

doi:10.1007/5584_2020_573

Cited by 6 publications

(13 citation statements)

References 24 publications

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“…Through this work, we have shown that both septicemia and PJI clinical isolates are stimulated to aggregate upon contact with BSF. These observations, along with previous studies demonstrating that synovial fluid-induced aggregation occurs with Gram-negative Enterobacter isolates, led us to conclude that a broad range of organisms display an aggregative phenotype ( 23 ).…”

Section: Discussionsupporting

confidence: 59%

“…Following staining, cells were washed twice with Ringer’s solution to remove excess dye before resuspension in either Ringer’s solution, 10% BSF in Ringer’s solution, 10% human serum in Ringer’s solution, or a combination of 5% BSF supernatant and 5% human serum in Ringer’s solution. Prior to addition, BSF was centrifuged for 1 min at 21,000 × g to separate out tissue components as previously described ( 23 ). BSF supernatant was used for all assays ( 23 ).…”

Section: Methodsmentioning

confidence: 99%

“…Prior to addition, BSF was centrifuged for 1 min at 21,000 × g to separate out tissue components as previously described ( 23 ). BSF supernatant was used for all assays ( 23 ). Cells were incubated in the respective host fluid treatments for 1 h, stagnantly at 22°C.…”

Section: Methodsmentioning

confidence: 99%

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Synovial Fluid-Induced Aggregation Occurs across Staphylococcus aureus Clinical Isolates and is Mechanistically Independent of Attached Biofilm Formation

et al. 2021

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Section: Discussionsupporting

confidence: 59%

Section: Methodsmentioning

confidence: 99%

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Synovial Fluid-Induced Aggregation Occurs across Staphylococcus aureus Clinical Isolates and is Mechanistically Independent of Attached Biofilm Formation

et al. 2021

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“…P. aeruginosa wild-type reference strain PAO1 was inoculated in 50% BSF/PBS for planktonic or suspended culture and lawn biofilm growth (13). BSF was used as the sole nutrient source and has been used as a model for bacterial growth in human synovial fluid (14)(15)(16)(17)(18).…”

Section: Pao1 Growth In 50% Bsf/pbsmentioning

confidence: 99%

“…The metabolic differences between the uninoculated BSF control and P. aeruginosa PAO1 in the suspended culture and static lawn biofilm phenotypes grown in BSF were identified by untargeted NMR-based metabolomics. For all samples, their 2D 13 C-1 H HSQC and 2D 1 H-1 H TOCSY spectra were processed and uploaded to the COLMARq web server for metabolite identification and quantification as previously described (8). A representative 13 C-1 H HSQC spectrum for each group (control, suspended, and biofilm) is shown as a color-coded overlay in Fig 1 . The spectra display a large number of distinct cross-peaks belonging to many detectable metabolites in each sample.…”

Section: Untargeted Metabolomics Analysis Of Bsf and P Aeruginosa In ...mentioning

confidence: 99%

Differential metabolism between biofilm and suspended Pseudomonas aeruginosa cultures in bovine synovial fluid by 2D NMR-based metabolomics

Leggett

Bruschweiler‐Li

et al. 2022

Preprint

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Total joint arthroplasty is a common surgical procedure resulting in improved quality of life; however, a leading cause of surgery failure is periprosthetic joint infection. Periprosthetic infection often involves biofilms, making treatment challenging. Periprosthetic joint infections are difficult to diagnose by traditional culturing methods and there are no clinical biomarkers for the presence of biofilms. Further, the metabolic state of pathogens in the joint space is difficult to diagnose, the mechanism of their tolerance to antibiotics and host defenses is not well understood, and their culturing in the laboratory is challenging. Thus, there is a critical need for improved pathogen- and phenotype-specific diagnosis as well as improved treatment strategies toward better patient outcomes. Here, we present a quantitative, untargeted NMR-based metabolomics strategy for Pseudomonas aeruginosa suspended culture and biofilm phenotypes grown in bovine synovial fluid. We identified 21 unique metabolites as potential markers of P. aeruginosa and one unique marker of the biofilm phenotype in synovial fluid. Significant differences in metabolic pathways were found between the suspended culture and biofilm phenotypes including creatine, glutathione, alanine, and choline metabolism and the tricarboxylic acid cycle. These unique metabolite and pathway differences have the potential to serve as targets for P. aeruginosa and specifically biofilm diagnosis and biofilm control in synovial fluid.Author SummaryJoint replacement surgery is a common procedure frequently required in later stages of life due to damage in the joint. Over one million joint replacement surgeries are performed annually with rates increasing every year. A devastating complication associated with joint replacement is the development of infection around the implant device in the joint space, known as a periprosthetic joint infection. Bacteria in the joint space can form a biofilm, which is a gel-like matrix encasing the cells that increases resistance to treatment and exacerbates chronic infections. A particular challenge for the diagnosis of biofilm-mediated periprosthetic joint infections is the slowly growing nature of biofilm-mediated phenotypes, resulting in frequent failure to detect these bacteria by clinical microbiological culturing methods. Small molecule metabolites are uniquely produced by strains of bacteria in the biofilm versus planktonic or suspended culture phenotype. Identification of metabolites as specific markers of infection and biofilm could allow a new culture-free diagnostic approach to diagnose infection by biofilm. Furthermore, knowledge of metabolic pathway populations in biofilm in joint fluid could point to specific targets to prevent biofilm formation in the joint space.

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The biofilm life cycle: expanding the conceptual model of biofilm formation

et al. 2022

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Bacterial biofilms are often defined as communities of surface attached bacteria. Biofilms are typically depicted with a classic mushroom-shaped structure that is a characteristic of Pseudomonas aeruginosa. However, it has become evident that this is not how all biofilms develop, especially in vivo and in the environment where biofilms often are observed as none surface attached aggregates. In this Review we describe the rationale behind the 5-step model and why it fails to capture many aspects of bacterial biofilm physiology, and aim to present an expanded developmental model for biofilm formation that is flexible enough to include all the diverse scenarios and microenvironments where biofilms are formed. With this new expanded inclusive model, we introduce a common platform for developing our understanding of biofilms and antibiofilm strategies that can be tailored to the microenvironment that is being investigated.

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Synovial Fluid Mediated Aggregation of Clinical Strains of Four Enterobacterial Species

Cited by 6 publications

References 24 publications

Synovial Fluid-Induced Aggregation Occurs across Staphylococcus aureus Clinical Isolates and is Mechanistically Independent of Attached Biofilm Formation

Synovial Fluid-Induced Aggregation Occurs across Staphylococcus aureus Clinical Isolates and is Mechanistically Independent of Attached Biofilm Formation

Differential metabolism between biofilm and suspended Pseudomonas aeruginosa cultures in bovine synovial fluid by 2D NMR-based metabolomics

The biofilm life cycle: expanding the conceptual model of biofilm formation

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