Presentation Title

Investigation of Marine Bacterial Compounds That Inhibit Biofilm Production in Mycobacteria

Faculty Mentor

Jacqueline A. Trischman

Start Date

17-11-2018 10:15 AM

End Date

17-11-2018 10:30 AM

Location

C335

Session

Oral 2

Type of Presentation

Oral Talk

Subject Area

physical_mathematical_sciences

Abstract

Tuberculosis (TB) kills more than one million people annually. Bacteria of the Mycobacterium genus, including M. tuberculosis, build a complex cell wall containing mycolic acids. This cell wall is difficult to penetrate, so specialized antibiotics are needed. Even with newly developed drugs, bacteria adapt quickly and exhibit resistance at an alarmingly rapid pace.One adaptation that allows the community to survive is production of biofilms. Formation of biofilm is one of many quorum sensing behaviors known in pathogenic mycobacteria. This additional layer surrounds a microenvironment where bacteria can thrive with a very low concentration of antibiotic. Thus, one alternative method to treat TB is to control biofilm formation.

In this research, a set of marine bacterial strains, including several bacteria that exhibited swarming behaviors and several from the same environmental samples that did not, were cultured, extracted, and analyzed by 1H NMR and LC-MS as well as in newly-developed biofilm and growth inhibition assays. Initial results showed one group of bacteria produced an organic compound that induced biofilm production in mycobacteria. This was an unexpected result. One representative strain producing a strong biofilm inducer was grown on large scale (10L) then extracted using progressively less polar eluents on a reversed-phase SPE column. The biofilm-inducing fraction was then separated using HPLC. One major component was analyzed spectroscopically using 1D and 2D NMR techniques along with Mass Spectrometry. This compound could result in a strategy to interfere with biofilm formation in mycobacteria, thus making antibiotics more effective.

Summary of research results to be presented

The results of a survey of growth and biofilm inhibiting activities in the extracts of 9 swarmers and 7 non-swarmer cultures will be reviewed. The set of 16 bacteria were isolated from the same sample of marine sediment and water. The bacteria were all grown on small-scale, extracted, then analyzed for chemical interest and biological activities. The strain UA350 produced a compound that was found in several of the strains tested. I will go through the grouping we saw in the chemistry and the surprising result that some of the extracts, including UA350, showed no growth inhibition, but significant biofilm induction.

UA350 was then grown by using a large-scale culture strategy and a scaled-up version of the same extraction method, resulting in three fractions. After chemical and biological analyses, the most polar fraction was separated using HPLC. This separation method development took a great deal of time, so strategies and results will be discussed thoroughly. A structural discussion of one of the main components of the extract will include analysis of 1D and 2D NMR and mass spectrometric data.

This document is currently not available here.

Share

COinS
 
Nov 17th, 10:15 AM Nov 17th, 10:30 AM

Investigation of Marine Bacterial Compounds That Inhibit Biofilm Production in Mycobacteria

C335

Tuberculosis (TB) kills more than one million people annually. Bacteria of the Mycobacterium genus, including M. tuberculosis, build a complex cell wall containing mycolic acids. This cell wall is difficult to penetrate, so specialized antibiotics are needed. Even with newly developed drugs, bacteria adapt quickly and exhibit resistance at an alarmingly rapid pace.One adaptation that allows the community to survive is production of biofilms. Formation of biofilm is one of many quorum sensing behaviors known in pathogenic mycobacteria. This additional layer surrounds a microenvironment where bacteria can thrive with a very low concentration of antibiotic. Thus, one alternative method to treat TB is to control biofilm formation.

In this research, a set of marine bacterial strains, including several bacteria that exhibited swarming behaviors and several from the same environmental samples that did not, were cultured, extracted, and analyzed by 1H NMR and LC-MS as well as in newly-developed biofilm and growth inhibition assays. Initial results showed one group of bacteria produced an organic compound that induced biofilm production in mycobacteria. This was an unexpected result. One representative strain producing a strong biofilm inducer was grown on large scale (10L) then extracted using progressively less polar eluents on a reversed-phase SPE column. The biofilm-inducing fraction was then separated using HPLC. One major component was analyzed spectroscopically using 1D and 2D NMR techniques along with Mass Spectrometry. This compound could result in a strategy to interfere with biofilm formation in mycobacteria, thus making antibiotics more effective.