A STUDY ON THE IN VITRO ANTIMICROBIAL ACTIVITY OF CEFTAZIDIME,

HONEY, AND CEFTAZIDIME PLUS HONEY VERSUS Pseudomonas

aeruginosa

Caballero, Rayzelle Anne

Cantela, Marra Monique
Lebrudo, Maria Janina
Morales, Karen Loura Mae
Pagtakhan, Ma. Nica Rielle
Principe, Larizze Leonica
Samson, Jennifer

MELECIA A. VELMONTE, M.D.

Research Adviser

April 23, 2015

 Chapter I

INTRODUCTION

BACKGROUND OF THE STUDY

Pseudomonas aeruginosa is member of the Gamma Proteobacteria

class of Bacteria. It is a Gram-negative, aerobic rod belonging to the bacterial

family Pseudomonadaceae. Since the revisionist taxonomy based on conserved

macromolecules (e.g. 16S ribosomal RNA) the family includes only members of

the genus Pseudomonas which are cleaved into eight groups. Pseudomonas

aeruginosa is the type species of its group. which contains 12 other members.

(Kenneth Todar, Ph.D.2012)

Like other members of the genus, Pseudomonas aeruginosa is a free-

living bacterium, commonly found in soil and water. However, it occurs regularly

on the surfaces of plants and occasionally on the surfaces of animals. Members

of the genus are well known to plant microbiologists because they are one of the

few groups of bacteria that are true pathogens of plants. In fact, Pseudomonas

aeruginosa is occasionally a pathogen of plants. However, Pseudomonas

aeruginosa has become increasingly recognized as an emerging opportunistic

pathogen of clinical relevance. Several different epidemiological studies track its

occurrence as a nosocomial pathogen and indicate that antibiotic resistance is

increasing in clinical isolates. (Kenneth Todar, Ph.D.2012)

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 Pseudomonas aeruginosa is an opportunistic pathogen, meaning that it

exploits some break in the host defenses to initiate an infection. In fact,

Pseudomonas aeruginosa is the epitome of an opportunistic pathogen of

humans. The bacterium almost never infects uncompromised tissues, yet there is

hardly any tissue that it cannot infect if the tissue defenses are compromised in

some manner. It causes urinary tract infections, respiratory system infections,

dermatitis, soft tissue infections, bacteremia, bone and joint infections,

gastrointestinal infections and a variety of systemic infections, particularly in

patients with severe burns and in cancer and AIDS patients who are

immunosuppressed. Pseudomonas aeruginosa infection is a serious problem in

patients hospitalized with cancer, cystic fibrosis, and burns. The case fatality rate

in these patients is near 50 percent. . (Kenneth Todar, Ph.D.2012)

. In vitro, ceftazidime is presently the most active cephalosporin available

against Pseudomonas aeruginosa. It is the antibiotic of choice because of the

high susceptibility of Pseudomonas to it. However, due to the high cost of

treatment, toxicity, and high burden of disease, alternative treatments are being

explored such as natural products and use of ancient remedies.

Honey has been used as an ancient remedy in the treatment of infected

wounds. It has also been widely studied and proven for its antimicrobial property.

A study by Allen, K.L., Hutchinson, G., Molan, P.C. shows that honey is effective

against MRSA, β-hemolytic streptococci, and Vancomycin-resistant enterococci.

However, this and most of the studies used Manuka Honey from New Zealand.

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The use of Philippine honey and its antimicrobial effect against MRSA has not

yet been fully established.

STATEMENT OF THE PROBLEM

Studies conducted shown that honey has antibacterial activity similar to

antibiotics, against test organisms and provides alternative therapy against

certain bacteria. However, the use of local honey and its antimicrobial effect

against Pseudomonas aeruginosa has not yet been fully established.

In response to this problem our study attempts to answer the following

research questions:

1. Does Philippine honey have in vitro antimicrobial activity against

Pseudomonas aeruginosa?

2. Does Philippine honey have minimum concentration to inhibit

Pseudomonas aeruginosa?

3. Does the in vitro antimicrobial activity of Philippine honey provide additive

effect to Ceftazidime in inhibiting Pseudomonas aeruginosa?

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OBJECTIVES OF THE STUDY

In this study, the researchers would like to:

1. To determine the in vitro antimicrobial activity of Philippine honey in

inhibiting Pseudomonas aeruginosa

2. To determine the minimum concentration of Philippine honey needed to

inhibit Pseudomonas aeruginosa; and

3. To determine if Philippine honey can provide additive effect to Ceftazidime

in inhibiting Pseudomonas aeruginosa

HYPOTHESES

 Null Hypothesis: Philippine honey has no in vitro antimicrobial activity

against Pseudomonas aeruginosa; and as an adjunct to Ceftazidime, it

has no effect in inhibiting Pseudomonas aeruginosa.

 Alternative Hypothesis: Philippine honey has in vitro antimicrobial

activity against Pseudomonas aeruginosa; and as an adjunct to

Ceftazidime, it has an additive effect in inhibiting Pseudomonas

aeruginosa.

SIGNIFICANCE OF THE STUDY

Pseudomonas aeruginosa infection is already known to have increasing

prevalence worldwide, and in the Philippines, a growing number of cases are

noted in both hospital and community. Since Ceftazimide is the first-line agent

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for treating Pseudomonas infections, there is high likelihood that antibiotic

resistance would eventually develop. In one study by Campeau and Patel (2014),

it was noted that combining honey (i.e. Manuka honey) and traditional antibiotics

(i.e. Gentamycin and Vancomycin), emergence and spread of traditional

antibiotic resistance may be impeded. However, there have been no published

researches regarding combination of Philippine honey to Ceftazidime.

Also, infection with Pseudomonas aeruginosa incurs high treatment cost

resulting to high morbidity and mortality outcome and vice versa. Therefore,

alternative treatments, including honey, are being widely explored due to cost

effectiveness and ability to impede antibiotic resistance.

In view of the significance of the study – A Study on the In Vitro

Antimicrobial Activity of Ceftazidime, Honey, and Ceftazidime Plus Honey Versus

Pseudomonas aeruginosa– It aims to determine the in vitro antimicrobial activity

of Philippine honey in inhibiting Pseudomonas aeruginosa and benefits the

following:

Academe/Researchers: This study will serve as future reference for studies

related to the determination of the antimicrobial effectiveness and activity of

honey.

Healthcare workers: This study would help them in providing an alternative

treatment for bacterial infections and in realizing the potential of honey as an aid

in decreasing the morbidity and mortality outcomes of Pseudomonas aeruginosa.

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Patients: This study will provide an alternative therapy that is cost effective

adjunct to traditional antibiotics which can help impede antibiotic resistance.

SCOPE AND LIMITATION

The study is limited to investigate the in vitro antimicrobial activity of

Philippine honey alone and as an adjunct to Ceftazidime in inhibiting

Pseudomonas infection. Its effectiveness in other forms of bacterial infections will

not be determined. Furthermore, the identification of Pseudomonas strain used

as the clinical isolate for the study was not done.

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 CONCEPTUAL FRAMEWORK

INPUT PROCESS OUTPUT

 Isolation of Anti microbial  Determination
pseudomona activity assay of minimum
s aeroginosa using a filter inhibitory
from paper disc concentration
wounded diffusion method  The In Vitro
individuals Antimicrobial
 Honey from : honey+ activity of
Department Ceftazidime in Ceftazidime,
of Agriculture each filter paper honey, and
of the disc honey filter Ceftazidime
University of paper disc,
the Ceftazidime filter
Philippines- paper disc,
Los Baños,
Laguna
 Preparation
of commercial
antibiotics