Antimicrobial activity of plant extracts from several genera

Joel Alicea Santa Dra. Jannette Gavilln PhD Heiselle Marquez, B.Sc.

Introduction
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Tramil is an association dedicated to scientifically validate the use of traditional medicinal plants for primary health care (APS). Their vision is to be the interdisciplinary program of reference in the identification, validation and diffusion of medical uses of plants that can impact public health. Tramil directs their studies tying them principally to applied scientific investigation based on the efficiency and security of medicinal plants.

Review: Herbal species

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Plantago major has been used as a treatment for renal infections caused by Escherichia coli, Staphylococcus aureus and Enterococcus spp. Cucurbita moschata is used to treat burns on various parts of the body. It is believed that this plant has antimicrobial action against Escherichia coli, Staphylococcus aureus and Enterococcus spp. Plectranthus amboinicus is used as an antimicrobial for Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus spp. It is also used as treatment for ear infections.

Problem
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Are plants traditionally used in Caribbean folk medicine effective treatments for ear infections, renal infections and burns? Can these extracts inhibit bacterial growth? If so, how significant is that inhibition?

Objectives
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Validate that Plantago major, Cucurbita moschata and Plectranthus amboinicus have antimicrobial effect against Escherichia coli, Staphylococcus aureus, Enterococcus spp and Pseudomonas aeruginosa . Determine the effectiveness of these plants for the treatment of ear infections, burns, renal infections. Give specific recommendations of the efficiency of these extracts for use as primary treatment for the health problems stated before.

Experiment

Plants Studied
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Plantago Major (leaves) : Used for renal infections in Dominica, Dominican Republic, Haiti, Guadeloupe, Martinique and Tobago. Cucurbita Moschata (leaves): Used for skin burns in Haiti and Dominican Republic Plectranthus amboinicus (leaves): Used for ear infections in Quintana Roo, Mxico.

Plants
Plantago Major Cucurbita Moschata Plectranthus amboinicus

://www.gardenorganic.org.uk/organicweeds/weed_information/weed.php

www.kuerbis.net/pflanze/bio/Kuerbis-Blaetter

http://pics.davesgarden.com/pics/pdb_Dinu

Bacteria
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Staphylococcus aureus, is present in the middle ear when there is an infections of the middle ear. Escherichia coli is a bacteria usually present in the large intestine and is responsible of 90% of renal infections. Enterococcus spp is also believed to cause renal infections. Pseudomonas aeruginosa is the cause of many infections that cause damage to the organism including the ear.

Methodology
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Plant specimens were recollected. A taxonomist verifies the plant specie An extract of each plant is prepared.
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Leaves were weighed. Cleaned with sterile water. Leaves were smashed. Leaves were cut in a blender. The mixture was filtered. The extracts were autoclaved to sterilize. Repeat for each plant.

Methodology
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The bacteria were inoculated in Luria-Bertani Broth (LB broth). For 24 hours. In an incubator at 36 C. Petri Plates were Prepared with Tryptic Soy Agar (TSA).

Methodology
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Petri dish preparation:

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Sterilize area with Lysol. Have an open flame to have a sterile area around the flame to work in an avoid outside contamination. Mark petri plates.

Metodologa
S. Aureus P.Aeruginosa E. Coli
Cucurbita Moschata Plantago Major

Enteroccus spp

Plectranthus amboinicus

N= 3

Methodology
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Bacteria were inoculated onto the petri plates using a spread method. Filter paper disks were soaked in the plant extracts. The filter paper disks were put on the center of each plate using the disk diffusion method. For the control plates the paper disks were soaked in sterile water. The petri plates with the bacteria were inoculated for 24 h at 36 Celcius. After this period the plates were removed from the incubator and the inhibition halo was measured for each plate.

Results
S. Aureus P.Aeruginosa E. Coli Enteroccus spp

Petri Plate

Cucurbita Moschata

+
+++ + ++ +++

Plantago Major

+ +

+ +

+ +

+ +

Plectranthus amboinicus

Disc radius= .2cm Light inhibition =.2-.5cm(+) Moderate inhibition=.5-1cm(++) Strong inhibition= >.1cm- (+++)

Primary Analysis of Results

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The antimicrobial activity of Plantago Major, Cucurbita Moschata, Plectranthus amboinicus resulted in a light inhibition of the growth of Staphylococcus aureus, Escherichia coli , Enterococcus spp, Pseudomonas aeruginosa. These results matched an experiment on Plantago Major extract done by Hoeletz that demonstrated that this plant has light antimicrobial activity against various bacteria's including E. Coli.

Future Work
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Future experiments should be done to conclude that the extracts from Plantago Major, Cucurbita Moschata and Plectranthus amboinicus do not have antimicrobial activity. The integration of other bacteria into the experimental design may result in different results. Changes in the methodology that can include specific amounts of extracts on each plate. The dilutions of these extracts and the addition of bacteria like Proteus Vulgaris and S. saprophyticus may give us more precise conclusions.

Conclusion
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After the completion of this part of the investigation the original hypothesis resulted to be negative. Our results suggest the extracts of Plantago Major, Cucurbita Moschata and Plectranthus amboinicus used in Caribbean folk medicine are not effective as primary treatments for burns, ear infections and renal infections caused by Staphylococcus aureus, Escherichia coli , Enterococcus spp and Pseudomonas aeruginosa. Further studies should be done to validate this conclusion.

References
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Hoeletz,Pessini,Sanches(2002), Screening of some plants used in the Brazilian Folk Medicine for the Treatment of Infectious Diseases.Mem Inst Oswaldo Cruz,Rio de Janeiro,Vol97(7) Hammer,Carson,Riley(1999), Antimicrobila activity of essential oils and other plant extracts. Journal of Applied Microbiology1999,85,985-990. Proestos,Chorianopoulos,Nychas(2005), RP-HPLC Analysis of thr Phenolic Compounds of plant Extracts. Invetigation of their Antioxidant Capacity and Antimicrobial Activity. J. Agric. Food Chem,2005,53.1190-1195.

Antimicrobial activity of plant extracts from several genera