1.

Introduction

Spondias pinnata is a deciduous tree

belonging to the family Anacardiaceae, widely

distributed in South-East Asian countries. It is

well-known as Common hog plum, Indian

mombin, Wild mango and Libas. The leaves

and fruits are used in culinary as sour in stews

and in filling fish dishes. The fruits are eaten

raw and can be made into jams and jellies. In

folklore, its bark, leaves and fruits are used in

the treatment of burns, sores and wounds.

Elsewhere, the bark is used for treatment of

diabetes. The wood of this tree is used for

various carpentry applications – mouldings,

interior finishing, turnery, carvings, etc.

Because of its lightness, it is suitable for the

manufacture of matchsticks, matchboxes,

boxes and crates.

Recent studies have proven the medicinal

uses of Libas. The ethyl acetate and

methanolic extracts of the plant’s stem

heartwood possess hepatoprotective activity

against carbon tetrachloride-induced

hepatotoxicity in rats. The hepatoprotective

activity could be due to the presence of

bioflavonoids. Study on chloroform and

methanol extracts of the bark of S. pinnata

produced significant diuretic and laxative

activity. Hypoglycemic activity was also

discovered through the study on adult Wistar

albino rats that showed the methanol extract

to have promising results comparable to

standard reference glibenclamide – an oral

hypoglycemic agent.

The objectives of this study are to determine

the antibacterial activity of Libas (Spondias

pinnata) leaves extract to Pseudomonas

aeroginosa and also to determine its degree

SENSITIVITY OF Pseudomonas aeroginosa TO LIBAS (Spondias pinnata) LEAVES EXTRACT

D. Antipuesto, C. De la Peña, J. Figueroa, A. Harder, A. Libo-on and J. TudilloUniversity of Negros Occidental – Recoletos

This scientific paper was an attempt to find out the antibacterial activity of Libas (Spondias pinnata) leaves extract to Pseudomonas aeroginosa and to determine its degree of sensitivity or resistance to the isolated bacteria in an appropriate range of the antimicrobial sample. In conducting the experiment on the antibacterial property of Libas, an extract was obtained on its leaves through concoction. Concoction is the process used to extract the dried and crushed Libas leaves. Then the sample was filtered, sterilized, evaporated and refrigerated. Then, sensitivity testing was done using the bacteria Pseudomonas aeroginosa in the prepared agar (Mueller- Hinton Base Agar). The plate was then divided into six sections with each section containing different samples. Streptomycin was being used for the positive control. The researchers used t-test dependent variable to detect the presence of the antibacteriocity property of the test substance. As the experiment has been conducted, the results showed that the Libas leaves extract shows positive result regarding its antibacterial property because of its greater zone of inhibition which is greater than that of the streptomycin. Thus, the data obtained in the experiment presents that the Libas leaves extract has an antibacteriocity property against Pseudomonas aeroginosa.

of sensitivity or resistance to the isolated

bacteria in an appropriate range of the

antimicrobial sample.

The significance of this study is to prove the

antibacterial activity of Libas using the sample

Pseudomonas aeroginosa. Sensitivity test was

used to show the degree of sensitivity or

resistance of the bacteria to the extracted

Libas leaves.

2. Review of Related Literature

B. Hazra, S. Biswas and N. Mandal.

Antioxidant and free radical scavenging

activity of Spondias pinnata

Dec. 9, 2008

Many diseases are associated with oxidative

stress caused by free radicals. Current

research is directed towards finding naturally-

occurring antioxidants of plant origin. The aim

of the present study was to evaluate the in

vitro antioxidant activities of Spondias

pinnata stem bark extract.

A 70% methanol extract of Spondias

pinnata stem bark was studied in vitro for

total antioxidant activity, for scavenging of

hydroxyl radicals, superoxide anions, nitric

oxide, hydrogen peroxide, peroxynitrite,

singlet oxygen and hypochlorous acid, and for

iron chelating capacity, reducing power, and

phenolic and flavonoid contents.

The extract showed total antioxidant activity

with a trolox equivalent antioxidant

concentration (TEAC) value of 0.78 ± 0.02. The

IC50 values for scavenging of free radicals were

112.18 ± 3.27 μg/ml, 13.46 ± 0.66 μg/ml and

24.48 ± 2.31 μg/ml for hydroxyl, superoxide

and nitric oxide, respectively. The IC50 for

hydrogen peroxide scavenging was 44.74 ±

25.61 mg/ml. For the peroxynitrite, singlet

oxygen and hypochlorous acid scavenging

activities the IC50 values were 716.32 ± 32.25

μg/ml, 58.07 ± 5.36 μg/ml and 127.99 ± 6.26

μg/ml, respectively. The extract was found to

be a potent iron chelator with IC50 = 66.54 ±

0.84 μg/ml. The reducing power was

increased with increasing amounts of extract.

The plant extract (100 mg) yielded 91.47 ±

0.004 mg/ml gallic acid-equivalent phenolic

content and 350.5 ± 0.004 mg/ml quercetin-

equivalent flavonoid content.

The present study provides evidence that a

70% methanol extract of Spondias

pinnata stem bark is a potential source of

natural antioxidants.

S. Mondal and G.K. Dash

Hypoglycemic activity of the bark of Spondias

pinnata Linn.kurz.

February 16, 2010

Diabetes, the most prevailing metabolic

disorder is attracting present research

attention towards it. In the present study, the

various extracts of the barks of Spondias

pinnata (Family: Rubiaceae) was evaluated for

hypoglycemic activity on adult Wistar albino

rats at dose levels of 300 mg/kg p.o. each

using normoglycaemic, glucose loaded and

alloxan induced hyperglycaemic rats.

Glibenclamide (2.5 mg/kg) was used as

reference standard for activity comparison.

Among the tested extracts, the methanol

extract was found to produce promising

results that is comparable to that of the

reference standard glibenclamide. The

preliminary phytochemical examination of the

methanol extract revealed presence of

flavonoids, tannins, saponins and terpenoids.

The present work justifies the use of the bark

in the folklore treatment in diabetes.

The results of the present study justify the

use of the barks of the plant for treating

diabetes as suggested in the folklore

remedies.

S. Mondal, G.K. Dash, S. Acharyya, D.K.

Brahma and S. Bal Studies on diuretic and

laxative activity of bark extracts of Spondias

pinnata February 16, 2010

The diuretic and laxative activity of different

extracts of the barks of Spondias pinnata were

studied in Wistar albino rats. Furosemide (10

mg/kg, p.o.) and agar-agar (300 mg/kg, p.o.)

were used as reference standards respectively

for activity comparison. The chloroform and

methanol extracts produced significant

diuretic and laxative activity. On the other

hand, the petroleum ether extract did not

reveal significant activity. Urinary levels of

sodium, potassium (by flame photometry) and

chloride (by titrimetry) were estimated.

Diuretics relieve pulmonary congestion and

peripheral edema and are useful in reducing

the syndrome of volume overload, including

orthopnea and paroxysmal nocturnal

dyspnoea. They decrease plasma volume and

subsequently venous return to the heart

(preload). This decreases cardiac workload,

oxygen demand and plasma volume, thus

decreasing blood pressure. Thus, diuretics

play an important role in hypertensive

patients. In present study, we can

demonstrate that the chloroform extract and

methanol extract of S. pinnata significantly

increased the urinary output as well as urinary

electrolyte concentration at a dose of 300

mg/kg, p.o. but the effect was found to be the

less potent in increasing the urinary output

when compared with the reference standard.

Further, the chloroform and methanol

extracts were found to be more effective in

enhancing urinary electrolyte concentration

for all the three ions tested (Na + , K + , Cl - ).

Petroleum ether extract on the other hand did

not increase urinary electrolyte

concentration. The increase in the ratio of

concentration of excreted sodium and

potassium ions indicates that the extracts

increase sodium ion excretion to a greater

extent than potassium, which is a very

essential requirement of an ideal diuretic with

lesser hypokalemic side effect.

The laxative activity study revealed

significant activity of the chloroform and

methanol extracts up to 8 h of drug

administration. The methanol extract was

found to be superior to that of the standard

drug and petroleum ether extract was found

to be least active.

Presence of phytoconstituents like

flavonoids, terpenoids, saponins, have been

previously found to be responsible for diuretic

and laxative activities in plants . The presence

of the said constituents in different extracts

of S. pinnata may be responsible for the

observed diuretic and laxative activities. The

exact mechanism exhibited by the extracts

can only be established after further

investigation.

3. Methodology

For this study, experimentation was used.

This type of devise was done to observe the

sensitivity of Pseudomonas aeroginosa to

Libas (Spondias pinnata) leaves extract. After

doing the experiment, the results would

determine if the actual outcome

corresponded to the anticipated result.

3.1 Materials

In conducting the experiment in

antibacterial activity of Libas, we have utilized

the following materials and apparatuses:

To accomplish the test sample to be tested

we used the following raw materials: leaves of

Libas, 100ml of ethyl acetone, Erlenmeyer

flask, volumetric flask, evaporating dish, vial,

glass cylinder, foil, and mortar and pestle.

For the sensitivity testing, the following

materials are used: Pseudomonas aeroginosa,

prepared agar, petri dishes,

micropippetor,and glass cylinder.

3.2 Preparation

In the Philippines, the leaves of different

plants are widely used as medication for

certain wounds or ailments. There are a lot of

leaves available in the country. The leaves are

most often used in culinary and folkloric. In

this study, Libas is used as a sample to test its

antibiotic properties; an extract was obtained

through concoction.

Concoction is the process used to obtain

the extract of Libas leaves by preparing 10

grams of Libas leaves per 100 ml of ethyl

acetone. Before extraction, we air-dried the

leaves and used mortar and pestle to crush it.

Upon obtaining the desired volume of sample

with ethyl acetone, itisthen filtered in a funnel

and sterilized gauze bandage. A 250ml

volumetric flask was used to serve as a

container for the extract. Then the extract

was evaporated until the desired volume is

obtained. After the evaporation process, the

extract is transferred in a vial and then

refrigerated.

3.3 Source of Microorganism

Pseudomonas aeroginosa, the

microorganism used in this experiment was

obtained by growing a pure culture courtesy

of Mr. Ronnie G. Gicana. Pseudomonas

aeroginosa is a common bacterium that can

cause disease in animals, including humans. It

is found in soil, water, skin flora, and most

man-made environments throughout the

world. It thrives not only in normal

atmospheres, but also in hypoxic

atmospheres, and has, thus, colonized many

natural and artificial environments. It uses a

wide range of organic material for food. For

an antimicrobial to be effective, it must reach

the site of infection in sufficient concentration

and duration to inhibit pathogenic growth.

3.4 Antibacteriocity (Sensitivity Testing)

To test the antibacterial properties of Libas,

sensitivity testing was used. Seven milliliter of

Mueller – Hinton base agar was poured into a

Petri dish. After the base agar has hardened, a

5 mL of top agar with 1mL ofPseudomonas

aeroginosa was added into the dish. Dividing

the plate into 6 sections and was labeled

appropriately as the samples. Seven glass

cylinders were placed on the top agar, six

cylinders for plant extracts and one for

streptomycin. Each glass cylinder contains

100µL of each plant extract and streptomycin

for the positive control. The result was

obtained after 24-48 hours of incubation at

room temperature. A positive result showed a

zone of inhibition around the glass cylinders.

4. Results and Discussion

This chapter presents, analyzes and

interprets the data gathered from the

experimentation of the Libas leaves extract.

The purpose of this study is to determine the

antibacteriocity of Libas leaves to

Pseudomonas aeroginosa. The following

statements will provide the data of the

experiment conducted to test the hypothesis

formulated in this study.

Table 1. The Antibacteriocity of Libas leaves

indicated by the measurements of its zone of

inhibition in millimeters against the positive

control antibiotic (Streptomycin) in Sensitivity

testing

Treatment Trail 1 Trial 2 Trial 3

Libas 21 21 24

Positive control 18 17 17

To prove the researchers hypothesis that

the zone of inhibition of exhibited by Libas

leaves extract coincides with the positive

control may indicate the presence of

antibacteriocity of the test substance, they

have used t-test dependent variable to

compare the means of the positive control

(Streptomycin) and the test substance (Libas

Leaves extract), if there is a significant

difference or none.

The table presented showed that there is a

significant difference between the positive

control and the test extract regarding their

zone of inhibition. It showed that the Libas

leaves extract zone inhibition in Pseudomonas

aeroginosa is greater than that of the

Sterptomycin. Thus the data obtained from

the experiment presents that Libas leaves

extract has an antibacteriocity property

against Pseudomonas aeroginosa.

5. Conclusion

Based on the results of the experiment

about the sensitivity of Pseudomonas

aeroginosa to Libas (Spondias pinnata) leaves

extract, we can say that the greater the zone

of inhibition of the test extract compare to

the positive control (streptomycin), the

greater its antibacteriocity property. In

conclusion to this study, the results gathered

from this experiment show that the Libas

leaves extract has an antibacteriocity property

greater than the positive control used in this

experiment.

6. References

1. http://www.stuartxchange.org/Libas.html

2. www.phcog.com

3. www.ncbi.nlm.nih.gov

4. S Mondal, GK Dash / Pharmacognosy

Magazine, 2009 | Volume : 5 | Issue : 19

5. S Mondal, GK Dash et al / Pharmacognosy

Magazine, 2009 | Volume : 5 | Issue : 19