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In Vitro Antimicrobial Activity of Pinus kesiya bark extract against
Staphylococcus aureus and Escherichia coliSECTION B
presentationoutline
I. INTRODUCTIONProblem and its backgroundConceptual FrameworkObjectiveStatement of the ProblemHypothesisSignificance of the StudyScope and Limitation
II. METHODOLOGYIII.RESULTS AND DISCUSSIONSIV.CONCLUSION and RECOMMENDATION
INTRODUCTION
Pine trees constitute a large genus
They are commercially important trees species
Carribean Pine, Pinus kesiya or Pinus insularis can be found in the highlands of Baguio City, Iba, Zambales and Tagaytay City in
the Philippines.
problem and its background
Escherichia coli and Staphylococcus aureus are common causes of Community Acquired
or Nosocomial Infections.
Escherichia coli is a true pathogenic organism and it exist with multiple genetic
subtypes. Kuhnert, 2000
Staphylococcus aureus an organism which can be found on human skin and frequently
inhabits the nose, and less commonly the throat.
problem and its background
INPUT (Literature search
finding about Pine tree, Escherichia coli, and
Staphylococcus aureus)
Plant collectionExtraction
Isolating pure cultures (streak plate technique)Agar well diffusion, MIC,
MBC
OUTPUTPinus kesiya bark
extract as an antimicrobial agent against E.coli and S.
aureus
conceptual framework
FEEDBACK
Peer Review
To determine if the Pinus kesiya bark extract has an antibacterial activity against Staphylococcus aureus and Escherichia coli.
objective
MAIN OBJECTIVE
SPECIFIC OBJECTIVES
objective
a. To determine if the active metabolites, tannins, flavonoids and terpenoids are present and can be isolated from the bark of the Pinus kesiya.
b. To determine if the isolated phytochemical compounds present in the crude extract of Pinus kesiya bark has an antibacterial activity against Staphylococcus aureus and Escherichia coli.
SPECIFIC OBJECTIVES
objective
c. To determine the lowest concentration of the extracts of Pinus kesiya bark that showed antibacterial activity against Staphylococcus aureus and Escherichia coli
d. To compare the antibacterial activity of the crude extract and the isolated compounds with the positive control.
SPECIFIC OBJECTIVES
objective
e. To determine if the crude extract and isolated compounds of Pinus kesiya bark has a bacteriostatic or bactericidal effect.
statement of the problem
Does the Pinus kesiya bark extract have an antibacterial
activity against Staphylococcus aureus and Escherichia coli?
Ho: The Pinus kesiya bark extract has no antibacterial activity against Staphylococcus aureus and Escherichia coli.
HI: The Pinus kesiya bark extract has an antibacterial activity against Staphylococcus aureus and Escherichia coli.
hypothesis
significance of the study
The determination of compounds with antibacterial activity against
clinically infectious strains of bacteria, while maximizing natural resources
found in the Philippines
significance of the study
Would help decrease the economic burden of medications and
the prevalence of diseases caused by the said organisms.
significance of the study
There are no previous studies about the antibacterial activity
of Pinus kesiya bark. Thus, this study will give baseline information in the use of Pinus kesiya as an antibacterial agent.
Antibacterial activity of the bark extract of Pinus kesiya acquired from Baguio City
limited to crude extraction and isolation of three phytochemical compounds present in the bark
scope and limitation
No purification and structural illucidation of the isolated phytochemical compounds were done.
No further characterization of the collected plant material and quantification of the isolated compounds through any molecular method were done.
scope and limitation
This study only determines the inhibitory effects of the extract against Escherichia coli and Staphylococcus aureus
scope and limitation
A comparative study was done between the succeptibility of the bacteria mentioned against the Crude extract, Isolated compounds and Positive Control.
No identification of the phytochemical compound that showed the greatest contribution in the antibacterial activity of the crude extract was done.
scope and limitation
METHODOLOGY
PROCEDURE
collection and
preparation of plant extracts
Microbial assay, MIC,
MBC of crude
extract
Isolation of active
compounds
Microbial assay, MIC,
MBC of isolated
compounds
1 2 3 4
ProcedurePlant collection and plant extraction
CRUDE ISOLATED COMPOUND
FLAVONOIDS TERPENOIDSTANNINS
Assay Assay Assay
MIC MIC MIC
MBC MBC MBC
Assay
MIC
MBC
RESULTS AND DISCUSSION
Pinus kesiya bark crude extract and the isolated compounds are
tested by using Agar-Well Method
Plant specimen verified and authenticated by Danilo N. Tandang of
Department of Botany, National Museum
plant material
750-gram sample of the bark was used in the experiment.
crude extract of Pinus kesiya
crude extract of Pinus kesiya
1. Tannins2. Flavonoids3. Terpenoids
isolated compounds
ISOLATED COMPOUNDSTANNINS
1• 400 grams of air-dried pine tree bark
2
• Isolated 2.7 grams (0.68%) of crude tannin
Qualitative test gave a blue-black color on Ferric Chloride test and definite precipitate in lead acetate test
ISOLATED COMPOUNDSFLAVONOIDS
1
• 3.02 grams flavonoids were isolated from the pine tree bark
2• Tested with Bate-Smith, Metcalf Method
Orange to strong red indicated the presence of flavonoids.
ISOLATED COMPOUNDSTERPENOIDS
1
• 2.19 grams (0.62%) of the product was obtained
2• Lieberman-Burchard Test
Blue to green and finally to purple color was observed.
ANTIMICROBIAL PROPERTY
MICROORGANISM POSITIVE-CONTROL
Staphylococcus aureus Oxacillin
Escherichia coli Ceftriaxone
Dimethyl sulphoxide or DMSO, was used as the solvent for the crude extract and was subsequently used as the Negative control.
POSITIVE CONTROL
NEGATIVE CONTROL
Crude Extract against S. aureus
Crude Extract against S. aureus
Crude Extract against E.coli
Crude Extract against E.coli
Minimum Inhibitory Concentration (MIC)
CRUDE EXTRACT
Minimum Bactericidal Concentration (MBC)
CRUDE EXTRACT
ANTIMICROBIAL PROPERTY OF ISOLATED COMPOUNDS
• Staphylococcus aureus is the only microorganism that showed susceptibility to the crude extract
Staphylococcus aureus
TANNIN EXTRACT
TANNIN EXTRACT vs. POSITIVE CONTROL
Minimum Inhibitory Concentration (MIC)
TANNIN EXTRACT
MIC of Tannin Extract against Staphylococcus aureus
Staphylococcus aureus
FLAVONOIDS EXTRACT
FLAVONOIDS EXTRACT vs. POSITIVE CONTROL
Staphylococcus aureus
TERPENOIDS EXTRACT
TERPENOIDS vs.POSITIVE CONTROL
Conclusion and Recommendations
Crude extract of Pinus Kesiya has an anti-bacterial activity against Staphylococcus aureus, and has no antibacterial activity
against Escherichia coli.
CONCLUSION
Terpenoids, flavonoids and tannins are present and isolated from the bark of
Pinus kesiya.
Only tannin extract has an antibacterial activity against S. aureus. however the
flavonoids and terpenoids did not show an antibacterial activity.
CONCLUSION
100 mg/ml was the lowest concentration of Pinus kesiya crude bark extract that showed antibacterial activity against
S. aureus.
CONCLUSION
Tannin extract did not show complete inhibition of S. aureus growth.
At 150 mg/ml concentration, Light growth was observed
CONCLUSION
Staphylococcus aureus showed susceptibility to tannins and was
resistant to flavonoids and terpenoids.
CONCLUSION
Crude extract is bacteriostatic and bactericidal against Staphylococcus aureus,
however, Tannin extract is neither bactericidal nor bacteriostatic.
CONCLUSION
RECOMMENDATIONS
For further research studies
1. determination of the susceptibility of other bacteria against the extract
2. different method of isolating the active compounds
3. purification of the isolated compounds
4. use a different solvent for the dilution of extracts.
RECOMMENDATIONS
Further characterization and quantification of phytochemical
compounds present in Pinus kesiya bark through molecular methods,
HPLC.
RECOMMENDATIONS
Determination of the active compound that shows the greatest contribution in the antibacterial activity of the crude
extract
THANK YOU!
In Vitro Antimicrobial Activity of Pinus kesiya bark extract against
Staphylococcus aureus and Escherichia coliSECTION B
HAPPY 25TH BIRTHDAY, JEREMY!