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Anti-virulent Disruption of Pathogenic Biofilms using Engineered Quorum-quenching Lactonases
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Microdialysis Sampling of Quorum Sensing Homoserine Lactones during Biofilm Formation
Alda Diaz Perez1, Kaleb Kougl1, Thaddeus W Vasicek1
1Department of Chemistry and Biochemistry , University of Arkansas , Fayetteville , Arkansas 72701 , United States.
Analytical Chemistry
|February 12, 2019
Summary
Researchers optimized microdialysis to collect bacterial quorum sensing molecules during biofilm formation. This method successfully sampled acylhomoserine lactones (AHLs) and acyl-oxohomoserine lactones (AOHLs) from V. harveyi biofilms.
Area of Science:
- Microbiology
- Analytical Chemistry
- Biotechnology
Background:
- Bacteria utilize quorum sensing (QS) for cell-to-cell communication, regulating group behaviors like biofilm formation.
- Quorum sensing molecules, such as acylhomoserine lactones (AHLs) and acyl-oxohomoserine lactones (AOHLs), are crucial for coordinating these processes.
- Understanding QS molecule dynamics during in situ biofilm development is essential for controlling bacterial communities.
Purpose of the Study:
- To optimize microdialysis sampling for direct collection of QS molecules during in situ biofilm formation.
- To quantify AHLs and AOHLs produced by *Vibrio harveyi* during early-stage biofilm development.
- To establish a novel method for real-time monitoring of QS signaling in complex bacterial communities.
Main Methods:
- Microdialysis probes with poly(ether sulfone) membranes were coated with fetal bovine serum to promote biofilm formation.
- Probes were used for in situ sampling of dialysates during and after *Vibrio harveyi* biofilm formation.
- Acylhomoserine lactones (AHLs) and acyl-oxohomoserine lactones (AOHLs) were extracted from dialysates using solid-phase extraction and quantified via liquid chromatography-mass spectrometry (LC-MS).
Main Results:
- Continuous sampling of five specific AHLs and AOHLs (C4-AHL, C6-AHL, C8-AHL, C6-OXO-AHL, C12-OXO-AHL) was achieved over a 4-day period.
- Detected concentrations of AHLs and AOHLs ranged from 1 to 100 ppb (ng/mL) and exhibited variability across sampling days.
- Successful collection of QS signaling molecules directly from the biofilm-forming surface was demonstrated.
Conclusions:
- Microdialysis sampling is a viable technique for collecting QS signaling molecules during in situ Gram-negative bacterial biofilm formation.
- This method provides a new avenue for studying the temporal dynamics of QS molecules in real-time.
- The optimized procedure offers potential applications in understanding and manipulating bacterial communication in various environments.
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