BIOFILM….

Dental caries…
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What is Biofilm?
 Biofilm is a community of
microorganisms that usually consist of
multiple species of bacteria that have the
ability to cause chronic pathogenic
infections in Humans.
 Biofilms are very difficult to eradicate with
conventional antimicrobial agents so they
create huge problems for practitioners.
 Bacterial biofilms have several potential
antimicrobial resistance mechanisms.
Drug Discovery Approach
Ethnomedicine Study
Eleutherine americana

Interview and discusson

with local healers

Clinical Isolate…..??
Drug From Plants Digitoxin
Digitalis Purpupea

Morphine
Papaver somniferum

Cocaine
Erythroxylum coca

Vinblastine
Catharantus Roseus

Paclitaxel (Taxol)
Taxus brevifolia
EFFECT OF PHENOLIC RICH EXTRACT AND FRACTION FROM Eleutherine
americana ON PLANKTONIC GROWTH AND BIOFILM FORMATION OF
Staphylococcus aureus

Dyke Gita Wirasisya1, Sudarsono2, Susi Iravati3

1Department of Pharmacy, Faculty of Medicine, Universitas Mataram, West Nusa Tenggara, Indonesia
2Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia
3Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia

Presented in : 1st International Conference on Global Health and Innovation

Mataram, 18–19th July 2019
 Research Question
1. Secondary metabolites identity of the sample (qualitative

and quantitative phytochemical analysis)

2. Effect on the planktonic growth (MIC and MBC)

3. Effect on biofilm formation (inhibition and degradation)

 01 Sample Preparation

RESEARCH • Plant was taken and cleaned from

METHODS the dirt.

• Plant freshly crushed and
macerated with ethanolic solvent.
• Fractionated using methanol.

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 02
Phytochemichal Screening
(qualitative)

• Assest using TLC and spray reagent to

dermine secondary metabolites
• Folin-ciocalteu  phenolic
03 TPC Determination
(quantitative)

• 2,4 DNPH  aldehyde/keton

Using spectropothometric
• 10% KOH  naftoquinone
method with Folin-
Ciocalteu reagent and
gallic acid as comparison.
 04
MIC and MBC determination

In Vitro macodillution
methods 05
BIOFILM TEST
• Microdillution with
crystal Violet staining.
• Inhibitory tested using
concentration below
MIC

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 RESULTS : Phytochemical Analysis

TPC (TOTAL PHENOLIC COMPOUND)

Extract  52,74 % GAE

Fraction  70,31 %GAE

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 Extract Fraction
No Bacteria
MIC MBC MIC MBC
1 ATCC 25923 0,50 mg/mL >4,00 mg/mL 0,50 mg/mL 2,00 mg/mL

2 SA 28 0,50 mg/mL >4,00 mg/mL 0,25 mg/mL >2,00 mg/mL

3 SA 913 1,00 mg/mL >4,00 mg/mL 0,50 mg/mL >2,00 mg/mL

4 SA 944 1,00 mg/mL >4,00 mg/mL 0,50 mg/mL >2,00 mg/mL

5 SA 947 1,00 mg/mL >4,00 mg/mL 0,50 mg/mL >2,00 mg/mL

6 SA 957 0,50 mg/mL >4,00 mg/mL 0,25 mg/mL >2,00 mg/mL

7 SA 758 1,00 mg/mL >4,00 mg/mL 0,25 mg/mL 2,00 mg/mL

Effect on the Planktonic Growth

(Fraction > Extract)

Its probably correlated with the TPC in the extract and fraction…
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 Biofilm Inhibition
No Bacteria
Extract Fraction BIOFILM
1 ATCC 25923 0,06 – 0,25 mg/mL 0,06 – 0,25 mg/mL
2 SA 28 0,06 mg/mL 0,06 – 0,12 mg/mL
3 SA 913 - -
4 SA 944 - 0,25 mg/mL
5 SA 947 0,12 – 0,25 mg/mL 0,25 mg/mL
6 SA 957 0,062 – 0,12 mg/mL 0,06 mg/mL
7 SA 758 0,062 – 0,12 mg/mL 0,06 – 0,12 mg/mL

Biofilm Degradation
No Bacteria
Extract Fraction
1 ATCC 25923 - -
2 SA 28 - -
3 SA 913 - -
4 SA 944 - 0,06 – 0,25 mg/mL
5 SA 947 - 0,06 – 0,25 mg/mL
6 SA 957 - -
7 SA 758 0,06 – 1 mg/mL 0,06 – 0,25 mg/mL
Tested below MIC concentration
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 CONCLUSION

1. Both extract and fraction showed secondary metabolites (phenolic, quinone

and aldehyde/ketone) with execption for sugar and its derivate only present

in extract.

2. The fraction (70,32 %GAE) have higher TPC than extract (52,74 % GAE)

3. The fraction have higher effect in planktonic growth of the bacteria.

4. Extract and fraction showed variable effect on inhibit and degrade biofilm

formation.

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Thank you!