ANTIBACTERIAL ACTIVITIES OF ALLIUM CEPA (ONION) ON TWO BACTERIAL ISOLATES

                                                      ABSTRACT

This study was conducted to ascertain the antibacterial activities of crude onion (Allium cepa) extract on some pathogenic bacteria isolates associated with ocular infection. This experimental study was carried out in the university premises, Michael Okpara University of Agriculture Umudike, Abia State, Nigeria. The antibacterial activity of onion extract on Staphylococcus aureus  and Escherichia coli was measured by disc diffusion method. Gentamycin was used as control for antibacterial activity. The effect on the antibacterial activities of Onion bulb (Allium cepa) was carried out and two different extracts were made from onion (Allium cepa) using distilled water. The extracts   include fresh onion extract and cold water extract. The antibacterial activity of the onion was tested on two bacteria isolates associated with ocular infections. The bacterial isolates include: Staphylococcus aureus, and Escherichia coli and. Among the two extracts, fresh onion extracts exhibited antagonistic effects on the test organisms ranging from observed zone of inhibition (average value) of 14mm for Staphylococcus aureus, 12mm for Escherichia coli. The antibiotic used as control was gentamycin with zone of inhibition of 17mm for Staphylococcus aureus, and 18mm for Escherichia coli. The minimum inhibitory concentration of the potent onion extracts shows positive inhibitory effect when an undiluted onion extracts was used. From the research conducted it was concluded that fresh onion extract could be the alternanative of the antibiotic to cure bacterial infection. However, pharmacological  Standardization and clinical assessment of this effect are essential before using onion extract as a preventive measure for human eye.

TABLE OF CONTENTS

Title                                                                                                         page

Title Page                                                                                                                                i

Certification                                                                                                                            ii

Dedication                                                                                                                              iii

Acknowledgement                                                                                                                  iv

Table of Content                                                                                                                     v

List of Tables                                                                                                                          vii                                                                                                                   

Abstract                                                                                                                                  viii

CHAPTER ONE

1.0        Introduction                                                                                                                1

1.2       Aim and Objectives                                                                                                    4

CHAPTER TWO

1.0           Literature Review                                                                                                       5

1.1           Botanical Source                                                                                                         5

1.2           Chemical Composition of Onion                                                                                6

2.3       Food Value of Onion                                                                                                                                                  7

2.4       Medicinal Uses of Onion                                                                                            7

2.5       Health Benefits Of Onion                                                                                           8

2.6       Medicinal Importance and Application                                                                      11

2.7       Antibacterial Activities of Onion                                                                               14

2.7.1    Antiviral Activities of Onion                                                                                      15

2.7.2    Antifungal Activities of Onion                                                                                   15

2.7.3     Antiparasitic Activity of Onion                                                                                 16

2.7.4     Antioxidant Activity of Onion                                                                                                                                                                                                                                                               16

2.7.5    Anticarcinogenic and Antimutagenic Activities of Onion                                      18

CHAPTER THREE

3.0       Materials and Methods                                                                                               20

3.1       Materials                                                                                                                     20

3.1.1    Sterilization Methods                                                                                                 20

3.1.2    Media Preparation                                                                                                      20

3.1.3    Preparation and Standardization of Bacterial Inoculum                                            20

3.2       Extraction of Crude from Onion Bulb (Allium cepa) of Onion                              21

3.2.1    Preparation of Concentration of Onion Extract                                                         21

3.3       Isolation, Identification and Confirmation of Some Pathogenic

 Bacteria Associated with Ocular Infection                                                               21

3.3.1    Gram Staining                                                                                                             21       

3.4       Determination of the Inhibitory Concentration of the Extracts of

The Isolates and to Measure the Zone of Inhibition                                                  22

3.4.1    Determination of Minimum Inhibitory Concentration of the Onion

            (Allium Cepa)                                                                                                             22

3.4.2    Determination of Minimum Bacterial Concentration (MBC)                                    22

3.5       Media                                                                                                                          23

3.6       Biochemical Reaction                                                                                               23

 CHAPTER FOUR

4.0       Results and Discussion                                                                                                26

CHAPTER FIVE

5.0       Discussion                                                                                                                   34

5.1       Conclusion                                                                                                                  35

5.2       Recommendation                                                                                                        35

REFERENCES

LIST OF TABLES

Table                                  Title                                     Page

4.1:     Identification and characterization of Bacterial Isolates                      28

4.2.1    Antimicrobial activity of Allium cepa (Onion) extracts on S. aureus 29

4.2.1    Antimicrobial activity of Allium cepa (Onion) extracts on E.coli                        30

4.3:      Percentage Occurrence of Bacteria Isolate (%)                                              31

4.4:       Minimum Inhibitory Concentration of Fresh and Cold

Water Onion Extract on Bacteria Isolate.                                           32

 4.5:     Maximum Bactericidal Concentration Of Fresh And Cold

Water Extract on Bacterial Isolates                                                    33

CHAPTER ONE

1.1   INTRODUCTION          

 Food borne disease is gradually becoming a serious risk to public health with the number of cases increasing yearly [Serthi S, Dutta A, Gupta B, Gupta S (2013)]. The cause is determined to be from bacteria, people become infected by either eating food infected with bacteria or intoxicated with enterotoxins produced from bacteria. Proper control of bacteria and effective sanitation will help to reduce the rate at which people become sick from eating food. Spices in general show both antibacterial and antioxidant characteristics, inhibiting the growth of bacteria and other pathogenic microbes [Serthi S, Dutta A, Gupta B, Gupta S (2013)]. Scientists in the past have conducted numerous experiments testing food borne pathogens with phytochemicals extracted from the spices [Erturk O (2006)]. The bacteria properties of active compounds extracted from spices and other plants in general based on recent findings have shown to affect the behavior of pathogenic bacteria and fungi in the agro food and medicinal sector (Serthi S, Dutta A, Gupta B, Gupta S (2013)].

 Onion has been revered throughout time not only for its culinary use, but also for its therapeutic properties. Consumption of onion is beneficial to human health as scientific studies show onion contains polyphenol molecules or phonutrients which includes flavonoids, tannins as well as allicin which possess antioxidant and antibacteria properties [Skrinjar M, Nemet N (2009)]. The main antibacterial agent in onion is quercetin and allicin (thio-2-propene-1-sulfuric acid-5-alkyl-esters), quercetin binds to the bacteria DNA gyrase while allicin inhibits certain thiolcontaining enzymes in the bacteria by the rapid reaction of thiosulfinates [Ankri S, Mirelman D (1999)].

This study investigates the antibacterial activity of photochemical extracted from onion (Allium cepa) against Staphylococcus aureus, Escherichia coli, Streptococci pneumonia using; Minimum inhibitory concentrations (MIC), Minimum bactericidal concentrations (MBC), and disc and agar well diffusion.

Allium cepa is usually thought of as a vegetable, It is also has a long history of medicinal use. Mainly the fleshy bulb that grows below the ground is used medicinally as well as for food but other parts of the plant also has a place in the traditional medicines.( K. P. Sampath, Kumar et al., 2010) Allium cepa is the common onion.

Onions are perennials that are cultivated for food worldwide. There are many varieties. Most onion bulbs are white, yellow, or red. The green stems and leaves are hollow and can reach 3 ft (1 m) in height. The plants bear small flowers that are usually white or purple. The fleshy bulb that grows below the ground is used medicinally as well as for food. Onion is believed to have a positive effect on the circulatory system. It has been used as a diuretic to reduce swelling. It is also thought to help reduce arteriosclerosis by lowering blood cholesterol levels and preventing the formation of blood clots. Onion has been used to treat diabetes and is reputed to lower blood sugar levels. Externally; fresh onion juice is used to prevent bacterial and fungal infections. It can be applied to wounds and stings on the skin, used to remove warts, used to stimulate hair growth, and even used to reduce unwanted skin blemishes. Warm onion juice dropped in the ear is said to help relieve earache. Baked onion is used to draw pus from abscesses.Modern scientific research supports many of the traditional uses for onion. Onion contains thiosulphinate, a compound that is effective in killing many common bacteria, including Salmonella typhi, Pseudomonas aeriginosa, and Escherichia coli. This finding supports the folk use of onion to treat wounds and skin infections and possibly its use for an upset stomach. Even more supportive are small clinical studies on humans that show that both fresh onions and commercial onion extracts actually lower blood cholesterol levels, lower blood pressure and help prevent the formation of blood clots. Although these studies have been done on only a small number of people, they are consistently supported by additional data from animal and test-tube studies.

The main properties of onion include antibacterial activity, cardiovascular support, hypoglycemic action, antioxidant/anticancer effect, and asthma protection. More research is needed in the area of asthma treatment, although certain compounds are said to inhibit bronchoconstriction. The bulb contains certain sulfur compounds that are known to be antibacteria.The onion also may be of benefit in cardiovascular disease, as it possesses hypolipidemic effects and has antiplatelet actions, retarding thrombosis. But certain lipid-reducing and blood pressure-lowering effects in humans have not yet been clinically proven. Some studies have been performed concerning diabetes treatment and ocular treatment by onion with promising results in animal and human experimentation. Although more research is needed on the use of onion as a treatment for diabetes in humans, many articles describe onion's benefits in improving glucose levels. The onion also is a proven antioxidant and may be helpful in treating certain cancers. More clinical research is needed to understand the many medicinal benefits of onion.

 It is usually consumed as fresh, however since losses of fresh onion in storage have been reported to be about 20-30% (Chadha and Sidhus, 1990), processed products are the most practical solution. Thus, the international market of onion is increasingly focused mainly on dehydrated products such as flakes, rings, granules, kibbles, powder, etc. and frozen or canned onions, or onion in vinegar, in brine or as essential oil, its commercial products being less abundant than those of garlic.

Dehydrated products have great commercial value not only by their culinary use but also their by medicinal properties as nutraceutical since they contains higher concentrations of beneficial compounds than the fresh forms (Lanzotti, 2006). In China, onion tea has long been recommended for several pathologies such as fever, headache, cholera and dysentery. Evidence from several investigations suggests that the biological and medical functions of onions are mainly due to their high content in organo-sulphur compounds (Augusti and Mathew, 1974; Wargovich et al., 1988). The primary sulphur-containing constituents in this vegetable are the S-alk(en)yl-L-cysteine sulphoxides. The isolation, identification and confirmation on some pathogenic bacteria isolates (Staphylococcus aureus, Escherichia coli, Staphylococcus pyogens, and Streptococci pneumonia ) associated with  ocular infection using crude onion extract and to determine the inhibitory concentration of onion extract of the isolate.

1.2       Aims and Objectives

The major aim of the study is to ascertain the antibacterial activity of crude onion extract on some pathogenic bacteria associated with ocular infection.

Specific Objectives of the Research

ü  To extract the antibacterial components of onion.

ü  To Isolate, identify and confirm some pathogenic bacteria associated with ocular infections.

ü  To determine the inhibitory concentration of the onion extracts of the isolates and to measure the zone of inhibition.

ü   The broad aim of this study is to ascertain the antibacterial activity of  crude onion extract  on some pathogenic bacteria associated with ocular infection.

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