Calcium carbonate mineralization is essential for biofilm formation and lung colonization

Cohen‐Cymberknoh, Malena; Kolodkin-Ga, Dror; Keren-Paz, Alona; Peretz, Shani; Brumfeld, Vlad; Kapishnikov, Sergey; Suissa, Ronit; Shteinberg, Michal; McLeod, Daniel; Maan, Harsh; Patrauchan, Marianna A.; Zamir, Gideon; Kerem, Eitan; Kolodkin‐Gal, Ilana

doi:10.1016/j.isci.2022.104234

Cited by 14 publications

(16 citation statements)

References 60 publications

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“…Ca 2+ is present in the body as a free ion concentration in the range of 1–5 mM. Such concentrations of calcium ions have been shown to form ionic bonds with alginate, thus producing a hydrogel-like biofilm structure with a diffuse arrangement of bacterial cells. ,, Ca 2+ also plays an important role in the structural integrity of a biofilm by electrostatically binding with eDNA to form bacterial aggregates . Thus, calcium-induced changes to EPS due to interactions with alginate and eDNA have been explored individually, ,− but the simultaneous bulk and microscale effects when both alginate and eDNA are prominent have not been well-characterized.…”

Section: Resultsmentioning

confidence: 99%

Physiological Concentrations of Calcium Interact with Alginate and Extracellular DNA in the Matrices of Pseudomonas aeruginosa Biofilms to Impede Phagocytosis by Neutrophils

Wells,

Currie,

Gordon

2023

Langmuir

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Biofilms are communities of interacting microbes embedded in a matrix of polymer, protein, and other materials. Biofilms develop distinct mechanical characteristics that depend on their predominant matrix components. These matrix components may be produced by microbes themselves or, for infections in vivo, incorporated from the host environment. Pseudomonas aeruginosa (P. aeruginosa) is a human pathogen that forms robust biofilms that extensively tolerate antibiotics and effectively evade clearance by the immune system. Two of the important bacterial-produced polymers in the matrices of P. aeruginosa biofilms are alginate and extracellular DNA (eDNA), both of which are anionic and therefore have the potential to interact electrostatically with cations. Many physiological sites of infection contain significant concentrations of the calcium ion (Ca2+). In this study, we investigate the structural and mechanical impacts of Ca2+ supplementation in alginate-dominated biofilms grown in vitro, and we evaluate the impact of targeted enzyme treatments on clearance by immune cells. We use multiple-particle tracking microrheology to evaluate the changes in biofilm viscoelasticity caused by treatment with alginate lyase or DNase I. For biofilms grown without Ca2+, we correlate a decrease in relative elasticity with increased phagocytic success. However, we find that growth with Ca2+ supplementation disrupts this correlation except in the case where both enzymes are applied. This suggests that the calcium cation may be impacting the microstructure of the biofilm in nontrivial ways. Indeed, confocal laser scanning fluorescence microscopy and scanning electron microscopy reveal unique Ca2+-dependent eDNA and alginate microstructures. Our results suggest that the presence of Ca2+ drives the formation of structurally and compositionally discrete microdomains within the biofilm through electrostatic interactions with the anionic matrix components eDNA and alginate. Further, we observe that these structures serve a protective function as the dissolution of both components is required to render biofilm bacteria vulnerable to phagocytosis by neutrophils.

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Section: Resultsmentioning

confidence: 99%

Physiological Concentrations of Calcium Interact with Alginate and Extracellular DNA in the Matrices of Pseudomonas aeruginosa Biofilms to Impede Phagocytosis by Neutrophils

Wells,

Currie,

Gordon

2023

Langmuir

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“…For mucoid biofilms grown with Ca 2+ , treatment with a combination of both alginate lyase and DNAse I is required to increase the phagocytic success of neutrophils Ca 2+ is present in the body as a free ion concentration in the range of 1-5mM. Such concentrations of calcium ions have been shown to form ionic bonds with alginate, thus producing a hydrogel-like biofilm structure with a diffuse arrangement of bacterial cells [8,43,44]. Ca 2+ also plays an important role in the structural integrity of a biofilm by electrostatically binding with eDNA to form bacterial aggregates [45].…”

Section: Resultsmentioning

confidence: 99%

Physiological concentrations of calcium interact with alginate and extracellular DNA in the matrices ofPseudomonas aeruginosabiofilms to impede phagocytosis by neutrophils

Wells,

Currie,

Gordon

2023

Preprint

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Biofilms are communities of interacting microbes embedded in a matrix of polymer, protein, and other materials. Biofilms develop distinct mechanical characteristics that depend on their predominant matrix components. These matrix components may be produced by microbes themselves or, for infections in vivo, incorporated from the host environment. Pseudomonas aeruginosa is a human pathogen that forms robust biofilms that extensively tolerate antibiotics and effectively evade clearance by the immune system. Two of the important bacterial-produced polymers in the matrices of P. aeruginosa biofilms are alginate and extracellular DNA (eDNA), both of which are anionic and therefore have the potential to interact electrostatically with cations. Many physiological sites of infection contain significant concentrations of the calcium ion (Ca2+). In this study we investigate the structural and mechanical impacts of Ca2+ supplementation in alginate-dominated biofilms grown in vitro and we evaluate the impact of targeted enzyme treatments on clearance by immune cells. We use multiple particle tracking microrheology to evaluate the changes in biofilm viscoelasticity caused by treatment with alginate lyase and/or DNAse I. For biofilms grown without Ca2+, we correlate a decrease in relative elasticity with increased phagocytic success. However, we find that growth with Ca2+ supplementation disrupts this correlation except in the case where both enzymes are applied. This suggests that the calcium cation may be impacting the microstructure of the biofilm in non-trivial ways. Indeed, confocal laser scanning fluorescence microscopy and scanning electron microscopy reveal unique Ca2+-dependent eDNA and alginate microstructures. Our results suggest that the presence of Ca2+ drives the formation of structurally and compositionally discrete microdomains within the biofilm through electrostatic interactions with the anionic matrix components eDNA and alginate. Further, we observe that these structures serve a protective function as the dissolution of both components is required to render biofilm bacteria vulnerable to phagocytosis by neutrophils.

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“…On the other hand, it has shown that bacterial biofilms contain inner mineral layer, contains calcite. Typically B. subtilis and P. aeruginosa make biofilms [36]. Chromate can interrelate with CaCO 3 in coprecipitated method via the MICP process, successfully removing chromium from Cr‐contaminated locations [37].…”

Section: Discussionmentioning

confidence: 99%

Microbial bio‐film calcite mediated removal of heavy metals from industrial wastewater of Kasur, Pakistan

2023

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Heavy metals in the industrial wastewater are an area of great concern as act as source of bioaccumulation in edible plants and posing a major health risk to humans like cancers. This study was planned by exploiting the bio‐film producing microbes that have the potential to remediate heavy metals by calcite mediated removal from industrial wastewater. Samples (n = 10) from a marble factory wastewater were collected. Samples were serially diluted and were spread on nutrient agar media supplemented with 2% urea and 0.28 g calcium chloride. All the isolates were observed for colony morphology, gram staining, and spore staining, for biochemical profile and for their efficacy in producing calcium carbonate crystals. All isolates showed cell densities at varying metal (chromium) concentrations ranging from 100 to 500 µg/mL. Determination of biofilm formation is performed by recording Optical density (OD = 600 nm). Normalized biofilm (570/600 nm) was formed. Different concentrations of chromium were used to measure their reduction ability and also by using tannery water. In tannery wastewater, significant reduction was recorded (p = 0.05) by AS4 bacterial isolate as compared to rest of the isolates and treatments. It showed remarkable chromium VI reduction ability.

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Calcium carbonate mineralization is essential for biofilm formation and lung colonization

Cited by 14 publications

References 60 publications

Physiological Concentrations of Calcium Interact with Alginate and Extracellular DNA in the Matrices of Pseudomonas aeruginosa Biofilms to Impede Phagocytosis by Neutrophils

Physiological Concentrations of Calcium Interact with Alginate and Extracellular DNA in the Matrices of Pseudomonas aeruginosa Biofilms to Impede Phagocytosis by Neutrophils

Physiological concentrations of calcium interact with alginate and extracellular DNA in the matrices ofPseudomonas aeruginosabiofilms to impede phagocytosis by neutrophils

Microbial bio‐film calcite mediated removal of heavy metals from industrial wastewater of Kasur, Pakistan

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