arbo , ma. adelle gia t. arteta , allen o. belmonte, pauline marie c
DESCRIPTION
Quorum Sensing Inhibition of Vibrio alginolyticus from Caulerpa lentillifera ( Lato ) on Pseudomonas aeruginosa. Arbo , Ma. Adelle Gia T. Arteta , Allen O. Belmonte, Pauline Marie C. . What is Quorum Sensing?. - PowerPoint PPT PresentationTRANSCRIPT
Quorum Sensing Inhibitionof Vibrio alginolyticus from Caulerpa lentillifera
(Lato) on Pseudomonas aeruginosa
Arbo, Ma. Adelle Gia T.Arteta, Allen O.
Belmonte, Pauline Marie C.
What is Quorum Sensing?
accumulation of signaling molecules enabling a single cell to sense the population density (Bhattacharya et al, 2008, pp. 30)
production and release of chemical signal molecules called autoinducers to coordinate gene expression
significant in biofilm formation and virulence of some microorganisms
an opportunistic bacterium that uses
quorum sensing in the formation of biofilms,
swarming motility, expolysaccharide
production, and cell aggregation.
(Bhattacharyya et al, 2008, pp. 30-33)
Pseudomonas aeruginosa
Figure 1. Pseudomonas aeruginosa(http://textbookofbacteriology.net/
pseudomonas.html)
Related Studies Givskov, Michael et al (2006)
-quorum sensing signaling systems of pathogens were used as central regulators for the expression of virulence factors
-Quorum sensing inhibitors - compounds are capable of reducing virulence and be used as treatment
Joint, Ian et al (2007) -the green seaweed Ulva has been shown to detect N-acylhomoserine lactones (AHLs)
Seaweeds suitable substrate
for epibiotic organisms because it releases a large amount of carbon into the surrounding environment
Figure 2. Seaweeds
http://davaoorientalinvest.com/main/?page_id=121
Caulerpa lentillifera• abundant in the
Philippines and cultivated as food and sold fresh and
salted
Figure 3. Caulerpa lentillifera
http://www.mesa.edu.au/marine_algae/gallery.asp)
Vibrio alginolyticus
a gram negative bacterium usually isolated from the surface of marine organisms such as seaweeds and stony corals
Some Vibrio species have quorum sensing inhibiting ability
Statement of the Problem
Main Problem:
Is Vibrio alginolyticus from seaweed, Caulerpa
lentillifera capable of inhibiting the quorum
sensing of Pseudomonas aeruginosa?
1. Are there zones of inhibition exhibited by Vibrio alginolyticus on Chromobacterium violaceum after agar overlay technique?
2. Are there zones of inhibition exhibited by Vibrio alginolyticus on Pseudomonas aeruginosa after agar overlay technique?
Specific Problems:
Significance of the Study
seaweed farmersantimicrobial agents economyPharmacology
Scope and Limitation
seaweed Caulerpa lentillifera use of indicator Chromobacterium violaceum 12472 use of lb broth
LimitationScope Quorum sensing inhibition assay antibacterial assay time observation time
Research Design
R X1 O1R X2 O2
Where:O1 & O2 = posttest observationX1 = quorum sensing inhibition assayX2 = antibacterial assay
Research Paradigm
Agar overlay of Pseudomonas aeruginosa
and Chromobacterium violaceum
Quorum sensing systems of
Pseudomonas aeruginosa;
Chromobacteriumviolaceum 12472, Vibrio
alginolyticus
Zones of inhibition of Pseudomonas
aeruginosa and
Chromobacterium violaceum 12472
Signaling molecules and growth conditions of the
bacteria
Independent Variables
Dependent Variables
Extraneous Variables
Linking Variables
Figure 4. Research Paradigm
Treatment/ General Procedure
Isolation of Vibrio alginolyticus
Figure 5. Taking samples of bacteria from Caulerpa
lentillifera
Figure 6. Agar plate inoculated with bacteria from Caulerpa
lentillifera
A. Quorum Sensing Inhibition AssayPreparation of Marine
agar plates and lb broth
Inoculation of Chromobacterium
violaceum 12472 and Vibrio alginolyticus on lb broth and marine agar,
respectively
24- Hour incubation
Overlay of lb broth on the marine agar plates inoculated
with Vibrio alginolyticus
24- Hour incubation
Observation and Data Gathering
Figure 7. Plate inoculated with C. violaceum and P. aeruginosa Figure 8. Inoculating lb
broth with C. violaceum
Figure 9. Test tube inoculated with C. violaceum
Figure 10. Overlay of lb broth on marine agar
after 24 hours.
Observation and Data Gathering
Preparation of marine agar plates and trypticase soy broth
Inoculation of Vibrio alginolyticus on marine agar
and of Pseudomonas aeruginosa on trypticase soy
agar
24-hour incubation
Overlay of trypticase soy on marine agar plates
24-hour incubation
B. Agar Overlay of Pseudomonas aeruginosa
Figure 11. TSB inoculated with Pseudomonas
aeruginosa
Figure 12 Marine agar inoculated with Vibrio
alginolyticus
Figure 13. Agar overlay of Pseudomonas aeruginosa
Figure 14. Overlay of TSB on marine agar after 24 hours
Results and DiscussionThere was no noticeable zone of inhibition
induced by Vibrio alginolyticus on both Chromobacterium violaceum and
Pseudomonas aeruginosa.
Figure 15. Marine agar plate of Vibrio alginolyticus after agar overlay of Chromobacterium
violaceum
Figure 16. Marine agar plate of Vibrio alginolyticus after agar overlay of Chromobacterium
violaceum
no verification that organisms had quorum sensing at that time mutations of organisms population of organisms conditions of bacterial growth
Further understanding of quorum sensing isolate more organisms exploit other techniques
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