REPUBLIC OF THE PHILIPPINES

DEPARTMENT OF EDUCATION
REGION VI- WESTERN VISAYAS
SCHOOLS DIVISION OF ILOILO
JANIUAY NATIONAL COMPREHENSIVE HIGHSCHOOL
Janiuay, Iloilo

EVALUATION OF THE INDIVIDUAL AND

COMBINATORY ANTIMICROBIAL ACTIVITY OF SAGE
(Salvia officinalis) AND ROSEMARY (Rosmarinus officinalis)
ESSENTIAL OIL EXTRACTS AGAINST Enterococcus
faecium, Proteus mirabilis, & Candida albicans

A Science Investigatory Project Proposal

Submitted in Partial Fulfilment of the Requirements in Research II
for Junior High School Special Science Curriculum

Submitted by:

HANS WEBSTER LABORDO

July 2017

NEMIA L. ALERTA
Adviser

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 CHAPTER I
INTRODUCTION

1.1 Background of the Study

Enterococci are a part of the normal intestinal flora of humans and animals. It has about
17 species and only two of those are disease causing microorganisms (Fraser, 2016). One
of them is Enterococcus faecium, a gram-positive bacterium that could be pathogenic and
causes neonatal meningitis, urinary tract infection (UTI) and in some cases, endocarditis –
a condition which can lead to stroke or heart failure (Kennedy, 2017; Kahn, 2016). Another
microorganism in this study is Proteus mirabilis, a gram-negative bacterium commonly
found in the intestinal tract. It causes more than 90% of all Proteus infections and causes
UTI and although less commonly, pneumonia (Gonzales, 2016). Pathogenic fungi are also
part of the intestinal human microbiota. Candida albicans is a species of yeast that infects
the vagina and the oral cavity, often resulting to vulvovaginal candidiasis or candida
vaginitis (Bennington-Castro, 2014).

As the antibiotic-resistant bacteria known as “superbugs” become more numerous and

more virulent, herbal medicine offers an alternative to increasingly ineffective drugs
(Buhner et al., 2014).

Rosemary is an herb that originally came from Southern Europe. Its oil and powder
are commonly used as spice and flavoring agent in food processing for its desirable flavor,
high antioxidant activity and lately as antifungal and antimicrobial agent (Lo et al., 2002;
Ouattara et al., 1997). According to the study of Moreno et al. (2006), rosemary plants are
rich sources of phenolic compounds with high antimicrobial activity against both gram-
positive and gram-negative bacteria commonly attributed to carnosic acid and carnosol. It
is clear that rosemary extracts have bioactive properties, but their antimicrobial activities
have not been deeply characterized.

As for sage, it is a native of the Mediterranean and have been traditionally used as a
remedy for common ailments like asthma, bronchitis, inflammations and depression

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(Ayotallahi et al., 2009; Rami et al., 2011; Walch et al., 2011; Khan et al., 2011). A number
of flavonoids and terpenoids are present in sage (Ayotallahi et al., 2009). Sage essential oil
is said to have carminative, antispasmodic, antiseptic and astringent properties. New
studies have shown that it is also effective in the treatment of a large range of diseases
including cardiovascular, nervous, digestive and pulmonary diseases among many others
(Loizzo et al., 2007; Radulescu et al., 2004).

Even with all these natural remedies, we are still largely dependent on synthetic drugs.
The result is that nowadays, multiple drug resistance has resulted from the indiscriminate
use of commercial antibiotics used in the treatment of infectious disease (Davis, 1994).

Hence there is a growing trend in screening new herbs with subsequent isolation of the
bioactive molecules from them. The traditional medicines have been derived from rich
traditions of ancient civilizations and heritage. Indigenous systems of medicine across the
world have enriched the present knowledge about the secondary metabolites and hence
much of the scientific investigations are associated and relay on traditional systems of
medicine (Parveen et al., 2011).

1.2 Statement of the Problem

Even with the rapidly advancing medical knowledge and the numerous discoveries in
this field, Filipinos have been known to rely on traditional practices and medication for
centuries especially in rural areas. Although rosemary and sage are relatively new to the
country, they are still a staple of traditional medicine.

Infections can be contracted from just about anywhere these days, even in a hospital.
So, I decided to use known medicinal herbs and screen them for pathogens that they have
not been screened for yet. It is in this rationale that rose (R. officinalis) and sage (S.
officinalis) are being tested for their antimicrobial properties against E. faecium, P.
mirabilis, C. albicans.

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1.3 Objectives of the Study

Generally, this study aims to evaluate the antimicrobial activities of Rosemary (R.
officinalis) and Sage (S. officinalis) essential oil extract.

Specifically, it aims to:

1. determine the individual antimicrobial activities of rosemary (R. officinalis) and

sage (S. officinalis) essential oil extract on E. faecium, P. mirabilis & C. albicans
based on the zone of inhibition (ZOI).
2. determine the combinatory antimicrobial activities of rosemary (R. officinalis) and
sage (S. officinalis) essential oil extract on E. faecium, P. mirabilis & C. albicans
based on the ZOI.
3. determine the best concentrations of rosemary (R. officinalis) and sage (S.
officinalis) essential oil extract based on the ZOI.

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REFERENCES:
Ayatollahi, A., Shojaii, A., Kobarfard, F., Mohammadzadeh, M. & Choudhary, M. (2009).
Two flavones from Salvia leriaefolia. Iran J Pharm, 8, 179-184.

Bennington-Castro, J. (2014). What is Candida Albicans? Retrieved from

http://www.everydayhealth.com/yeast-infection/guide/causes/candida-albicans/

Buhner, S. H. (2014). Herbal Antibiotics: An Effective Defense Against Drug-Resistant

‘Superbugs’. Mother Earth News. Retrieved from
http://www.motherearthnews.com/natural-health/herbal-remedies/herbal-
antibiotics-defense-zm0z13djzsor.aspx

Fraser, S. L. (2016). Enterococcal Infections. Retrieved from

http://emedicine.medscape.com/article/216993-overview

Gonzales, G. (2016). Proteus Infections. Retrieved from

http://emedicine.medscape.com/article/226434-overview#a4

Kahn, A. (2016). Endocarditis. Retrieved from

http://www.healthline.com/health/endocarditis#overview1

Kennedy, R. (2017). Enterococcus. Retrieved from http://www.medical-

library.net/content/view/523/41/

Khan, A., Najeeb-ur- Rahman & Alkharfy, G. S. (2011) Antidiarrheal and antispasmodic
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Lo, A. H., Liang, Y. C., Lin-Shiau, S. Y., Ho, C. T. & Lin, J. K. (2002). Carnosol, an
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Loizzo, M. R., Tundis, R., Menichini, F., Saab, A.M., Statti, G. A. et al. (2007) Cytotoxic
activity of essential oils from labiatae and lauraceae families against in vitro human
tumor models. Anticancer Res. 27, 3293-3299.

Moreno, S., Scheyer, T., Romano, C. S., Vojnov, A. A. (2006). Antioxidant and
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Ouattara, B., Simard, R. E., Holley, R. A., Piette, G. J. P. & Begin, A. (1997). Antibacterial
activity of selected fatty acids and essential oils against six meat spoilage organisms.
Int. J. Food Microbiol., 37, 155-162.

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Radulescu, V. & Chiliment, S. O. E. (2004). Capillary gas chromatography-mass
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