Recent advances in experimental design and data analysis to characterize prokaryotic motility

Dubay, Megan Marie; Acres, Jacqueline M.; Riekeles, Max; Nadeau, Jay

doi:10.1016/j.mimet.2022.106658

Cited by 15 publications

(8 citation statements)

References 96 publications

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“…Using Trackmate, a thresholding detector was applied with the intensity adjusted from 30 to 40 to define individual microspheres. Once all microspheres in all frames were detectable, we then utilized the simple LAP tracker, the tool specific for non-branching tracks, to quantify the lateral (XY) trajectories and the mean velocity of the microspheres ( Tsvirkun et al, 2017 ; Dubay et al, 2023 ). The plugin compiled all successive positions, constructed the track list, and exported data to an excel file for further analysis.…”

Section: Methodsmentioning

confidence: 99%

Development of a tomato xylem-mimicking microfluidic system to study Ralstonia pseudosolanacearum biofilm formation

Chu,

Laxman,

Abdelhamed

et al. 2024

Front. Bioeng. Biotechnol.

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The bacterial wilt pathogen Ralstonia pseudosolanacearum (Rps) colonizes plant xylem vessels and blocks the flow of xylem sap by its biofilm (comprising of bacterial cells and extracellular material), resulting in devastating wilt disease across many economically important host plants including tomatoes. The technical challenges of imaging the xylem environment, along with the use of artificial cell culture plates and media in existing in vitro systems, limit the understanding of Rps biofilm formation and its infection dynamics. In this study, we designed and built a microfluidic system that mimicked the physical and chemical conditions of the tomato xylem vessels, and allowed us to dissect Rps responses to different xylem-like conditions. The system, incorporating functional surface coatings of carboxymethyl cellulose-dopamine, provided a bioactive environment that significantly enhanced Rps attachment and biofilm formation in the presence of tomato xylem sap. Using computational approaches, we confirmed that Rps experienced linear increasing drag forces in xylem-mimicking channels at higher flow rates. Consistently, attachment and biofilm assays conducted in our microfluidic system revealed that both seeding time and flow rates were critical for bacterial adhesion to surface and biofilm formation inside the channels. These findings provided insights into the Rps attachment and biofilm formation processes, contributing to a better understanding of plant-pathogen interactions during wilt disease development.

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Section: Methodsmentioning

confidence: 99%

Development of a tomato xylem-mimicking microfluidic system to study Ralstonia pseudosolanacearum biofilm formation

Chu,

Laxman,

Abdelhamed

et al. 2024

Front. Bioeng. Biotechnol.

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“…It took 18 hours of incubation at 35°C, or until noticeable growth appeared. [22] Nitrate reduction test: 0.5 ml of nitrate broth was added in a clean test tube, was autoclaved for 15 minutes at 15 lbs pressure and 121°C, and was let to cool to room temperature. The tube was inoculated with a heavy inoculum of fresh bacterial culture and was incubated at 35°C for 2 hours.…”

Section: Gram Stainmentioning

confidence: 99%

Exhibition of <em>Corallopyronin A </em>an Antibiotic from Different Soil Environments in Egypt

Kassab

2024

Preprint

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Background: Across the world, antibiotic resistance is a critical challenge. To address such a problem, it has to be done to investigate new antibiotic sources. Aim of the study: Study of the antibacterial activity of Corallopyronin A in preclinical animal testing and randomized human clinical trials stages 1/2, as well as investigating the purification of Corallopyronin A from different soil conditions in Egypt. Type of the study: Screening experimental study. Methodology: Egypt's various soil conditions were tested to develop bacterial isolates that produced the antibiotic compound Corallopyronin A. Reversed phase HPLC was used to purify Myxopyronin A. The broth microdilution method and the paper disc diffusion assay determined the test antibiotic's minimum inhibitory concentration( MIC) and in vitro antibacterial activity. Moreover, in vivo antibacterial spectrum, adverse medication responses, and pharmacokinetics were found in phases 1/2 of randomized clinical studies involving humans and animal models. Results: Corallopyronin A was generated from the culture supernatant of the soil bacterial isolate Corallococcus coralloides M2, cultivated on a Casein yeast peptone( CYP) plate. The test antibiotic prevented the growth of several Gram-ve bacteria, including Escherichia coli, at MICs higher than 100 mcg/ ml; while blocking the development of many Gram +ve bacteria, with MICs ranging from 3 to 15 mcg/ ml. However, eukaryotic cells—such as those found in fungi and humans—were unaffected. A bactericidal consequence of the test antibiotic was detected in the inhibition of prokaryotic DNA-dependent RNA polymerase( RNLP). Conclusion: The synthesis of the bactericidal antibiotic Corallopyronin A from Corallococcus coralloides M2 isolated from different soil settings in Egypt intrigued the current investigation.

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“…It took 18 hours of incubation at 35°C, or until evident growth materialized. [22] Nitrate reduction test: 0.5 ml of Nitrate broth situated in a clean test tube was autoclaved for 15 minutes at 15 lbs pressure and 121°C, and was all-awed to cool to room temperature. The tube was inoculated with a heavy inoculum of unspoiled bacterial culture and was incubated at 35°C for 2 hours.…”

Section: Introductionmentioning

confidence: 99%

Screening and the Industry of <em>Myxopyronin B</em> Antibiotic from Different Soil Environments in Egypt

Kassab

2024

Preprint

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Background: Across the world, antibiotic resistance is a grave problem. To address this issue, it is necessary to investigate new antibiotic sources. Aim of the study: Egypt's diverse soil habitats were used to produce bacterial Myxopyronin B, which was then tested for antimicrobial activity in preclinical animal studies and randomized human clinical trials stages 1/2. Type of the study: Screening experimental study. Methodology: Egypt's various soil conditions were examined for the development of bacterial isolates that produced the antibiotic compound Myxopyronin B. Reversed phase HPLC was used to purify Myxopyronin B. To determine the test antibiotic's minimum inhibitory concentration( MIC) and invitro antibacterial activity, the Broth microdilution method and the Paper disc diffusion assay were utilized. Moreover, in vivo antibacterial spectrum, adverse medication responses, and pharmacokinetics were found in preclinical animal testing stages and phases 1/2 of randomized clinical studies involving volunteer humans. Results: Myxopyronin B was generated from the culture supernatant of Myxococcus SDU36, the main soil bacterial isolate cultured on a Casein yeast peptone plate. With MICs less than 100 mcg/ml, the test antibiotic prevented the growth of several Gram +ve bacteria, whereas, at MICs higher than 100 mcg/ml, it inhibited the development of a small number of Gram -ve bacteria, including Escherichia coli. However, eukaryotic cells—such as those found in fungi and humans—were unaffected. The test antibiotic was seen to inhibit prokaryotic DNA-dependent RNA polymerase( RNLP), indicating its bactericidal activity. Mean Cmax was 9-10 mcg/ ml at mean Tmax 1 hour when 600 mg dose was orally administered in randomized human clinical trials phases 1/2, and T1/2 reached 2.25 hours following first-order kinetics of elimination. Duration of its action was nearly 8 hours after oral administration. Rare toxicity was detected during preclinical and randomized human clinical trials phases 1/2 in the form of mild diarrhea and GI upset in less than 7 % of experimental candidates. It exhibited approximately 90% plasma protein binding, especially with albumin which resulted in prolonged therapeutic action. It showed a concentration dependant antibiotic killing effect. With the concentration-dependent killing pattern, the higher the drug concentration relative to the pathogen minimum inhibitory concentration( MIC), the greater the rate and extent of antimicrobial activity. Conclusion: The present study was promising due to the production of the bactericidal antibiotic Myxopyronin B from Myxococcus sp. SDU36 isolated from different soil environments in Egypt.

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Recent advances in experimental design and data analysis to characterize prokaryotic motility

Cited by 15 publications

References 96 publications

Development of a tomato xylem-mimicking microfluidic system to study Ralstonia pseudosolanacearum biofilm formation

Development of a tomato xylem-mimicking microfluidic system to study Ralstonia pseudosolanacearum biofilm formation

Exhibition of <em>Corallopyronin A </em>an Antibiotic from Different Soil Environments in Egypt

Screening and the Industry of <em>Myxopyronin B</em> Antibiotic from Different Soil Environments in Egypt

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