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

D. Antipuesto, C. De la Pea, J. Figueroa, A. Harder, A. Libo-on and J. Tudillo

University 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 (MuellerHinton 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 ttest 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.

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 plants 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 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.

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 g/ml, 13.46 0.66 g/ml and 24.48 2.31 g/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 g/ml, 58.07 5.36 g/ml and 127.99 6.26

2. Review of Related Literature

g/ml, respectively. The extract was found to be a potent iron chelator with IC50 = 66.54

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 naturallyoccurring antioxidants of plant origin. The aim of the present study was to evaluate the in vitro antioxidant activities of Spondias

0.84

g/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 quercetinequivalent flavonoid content. The present study provides evidence that a 70% methanol extract of Spondias

pinnata stem bark is a potential source of natural antioxidants.

pinnata stem bark extract. A 70% methanol extract of Spondias 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

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

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.

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

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

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

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 remedies. as suggested in the folklore

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.

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

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

diuretic and laxative activity. On the other

lesser The

hypokalemic laxative activity

side study

effect. revealed

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,

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 Presence be of least phytoconstituents active. like

micropippetor,and glass cylinder.

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 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 3. Methodology concoction.

investigation.

Concoction is the process used to obtain the extract of Libas leaves by preparing 10

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

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

corresponded to the anticipated result.

3.1 Materials In conducting the experiment in

was evaporated until the desired volume is obtained. After the evaporation process, the extract is transferred in a vial and then refrigerated.

antibacterial activity of Libas, we have utilized the following materials and apparatuses:

3.3 Source of Microorganism Pseudomonas aeroginosa, the

room temperature. A positive result showed a zone of inhibition around the glass cylinders.

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 but not only also in normal This chapter presents, analyzes and 4. Results and Discussion

interprets the data gathered from the experimentation of the Libas leaves extract. The purpose of this study is to determine the antibacteriocity Pseudomonas of Libas leaves The to

atmospheres,

in hypoxic

aeroginosa.

following

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.

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

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 100L of each plant extract and streptomycin for the positive control. The result was obtained after 24-48 hours of incubation at

control antibiotic (Streptomycin) in Sensitivity testing Treatment Libas Positive control Trail 1 21 18 Trial 2 21 17 Trial 3 24 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