Sensing Inhibition of Vibrio alginolyticus from Caulerpa lentillifera Lato on Pseudomonas aeruginosa Arbo Ma Adelle Gia T Arteta Allen O Belmonte Pauline Marie C ID: 555806
Download Presentation The PPT/PDF document "Quorum" is the property of its rightful owner. Permission is granted to download and print the materials on this web site for personal, non-commercial use only, and to display it on your personal computer provided you do not modify the materials and that you retain all copyright notices contained in the materials. By downloading content from our website, you accept the terms of this agreement.
Slide1
Quorum Sensing Inhibitionof Vibrio alginolyticus from Caulerpa lentillifera (Lato) on Pseudomonas aeruginosa
Arbo
, Ma.
Adelle
Gia
T.
Arteta
, Allen O.
Belmonte, Pauline Marie C. Slide2
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 microorganismsSlide3
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 aeruginosaFigure 1. Pseudomonas aeruginosa
(http://textbookofbacteriology.net/pseudomonas.html)Slide4
Related Studies Givskov, Michael et al (2006)quorum sensing signaling systems of pathogens were used as central regulators for the expression of virulence factorsQuorum sensing inhibitors - compounds are capable of reducing
virulence and be used as treatmentSlide5
Joint, Ian et al (2007) the green seaweed Ulva has been shown to detect N-acylhomoserine lactones (AHLs) Slide6
Seaweeds suitable substrate for epibiotic organisms because it releases a large amount of carbon into the surrounding environmentFigure 2. Seaweeds
http://davaoorientalinvest.com/main/?page_id=121Slide7
Caulerpa lentilliferaabundant in the Philippines and cultivated as food and sold fresh and saltedFigure 3. Caulerpa lentilliferahttp://www.mesa.edu.au/marine_algae/gallery.asp
)Slide8
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 abilitySlide9
Statement of the ProblemMain Problem:Is Vibrio alginolyticus from seaweed, Caulerpa lentillifera capable of inhibiting the quorum sensing of Pseudomonas aeruginosa? Slide10
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:Slide11
Significance of the Studyseaweed farmersantimicrobial agents economyPharmacologySlide12
Scope and Limitation seaweed Caulerpa lentillifera use of indicator Chromobacterium violaceum 12472 use of lb brothLimitationScope
Quorum sensing inhibition assay
antibacterial assay
time
observation timeSlide13
Research DesignR X1 O1R
X2 O2
Where:
O1
& O2 =
posttest
observation
X1
= quorum sensing
inhibition
assay
X2
= antibacterial assaySlide14
Research ParadigmAgar overlay of Pseudomonas aeruginosa and Chromobacterium violaceum
Quorum sensing systems of
Pseudomonas aeruginosa;
Chromobacteriumviolaceum
12472, Vibrio alginolyticus
Zones of inhibition of
Pseudomonas aeruginos
a and
Chromobacterium violaceum 12472
Signaling molecules and growth conditions of the bacteria
Independent Variables
Dependent Variables
Extraneous Variables
Linking Variables
Figure 4. Research ParadigmSlide15
Treatment/ General ProcedureIsolation of Vibrio alginolyticus
Figure 5. Taking samples of bacteria from
Caulerpa lentillifera
Figure 6. Agar plate inoculated with bacteria from
Caulerpa lentillifera
Slide16
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.Slide17
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 hoursSlide18
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 violaceumSlide19
no verification that organisms had quorum sensing at that time mutations of organisms population of organisms conditions of bacterial growthSlide20
Further understanding of quorum sensing isolate more organisms exploit other techniquesRecommendationsSlide21
Let us take this vital step together! Slide22
Thank you for listening!