ANTIMICROBIAL ACTIVITY OF CAPSICUM ANNUUM (CHILI PEPPER) EXTRACT ON PATHOGENIC MICROORGANISMS

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ABSTRACT

Extracts of Capsicum annuum were studied for their antimicrobial potency against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Shigella spp and Candida albicans using the disc diffusion technique. Qualitative and quantitative phytochemical screening of the ethanol and aqueous extracts of the plant showed the presence of alkaloids, flavonoids, saponins, tannins, steroids, cyanogens and phenols with alkaloids being the dominant compound with a percentage composition of 0.52%. Both the aqueous and ethanolic extracts of Capsicum annuum showed antimicrobial activity against all the test organisms. In aqueous extract, Capsicum annuum showed good inhibitory activity mainly against Escherichia coli and Shigella  spp with zone of inhibition of 19.67 mm ±1.16b and 14.33 mm ±1.52b respectively. The ethanol extract of the plant performed better against Staphylococcus aureus and Pseudomonas aeruginosa with zone of inhibition of 20.33 mm ± 3.06b and 14.33 mm± 0.58b respectively. Based on this finding, this extract may be an alternative to chemical preservatives and used as natural antimicrobial preservatives to reclaim the shelf-life of food.

 



TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgement                                                                                                                  iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          viii

List of Figures                                                                                                                         ix

Abstract                                                                                                                                  x


CHAPTER ONE

Introduction                                                                                                                            1

1.1       Background of the Study                                                                                            1

1.2       Objectives of the Study                                                                                              4


CHAPTER TWO

Literature Review                                                                                                                   5

2.1       Description of Capsicum annum (Chilli Pepper)                                                       5

2.2       The Origin of Chilli Pepper                                                                                        5

2.3      Varieties of Pepper                                                                                                    5

2.4        Intensity of Pepper                                                                                           7

2.5       Characteristics of Chilli Pepper                                                                                  7

2.6       Classification of Chilli Pepper                                                                                   8

2.7       Uses of Pepper                                                                                                            8

2.7.1    Culinary Uses                                                                                                             8

2.7.2    Medicinal Uses                                                                                                           9

2.7.3    Nutritional Uses                                                                                                          9

2.7.4    Crop Defense Uses                                                                                                     10

CHAPTER THREE

Materials and Methods                                                                                                           11

3.1       Collection of Materials                                                                                               11

3.2       Preparation of Samples                                                                                               11

3.3       Media Preparation                                                                                                      11

3.4       Production of Extract                                                                                                 12

3.5       Preliminary Phytochemical Screening                                                                       12

3.5.1    Qualitative Analysis of Phytochemicals                                                                    13

3.5.1.1  Test For Tannins                                                                                                         13

3.5.1.2     Test For Saponins                                                                                                   13

3.5.1.3    Test For Flavonoids                                                                                                 13

3.5.1.4    Test For Alkaloids                                                                                                   14

3.5.1.5    Test For Steroids                                                                                                      14

3.5.1.6    Test For Phenol                                                                                                        15

3.5.2      Quantitative Determination of the Phytochemicals                                                    15

3.5.2.1 Determination of Alkaloids                                                                                       15

3.5.2.2  Determination of Flavonoids                                                                                      16

3.5.2.3  Determination of Tannins                                                                                           16

3.5.2.4  Determination of Saponin                                                                                          18

3.5.2.5  Determination of Phenols                                                                                           19

3.5.2.6  Determination of Cyanogenic Glycoside (HCN)                                                       20

3.6       Antimicrobial Activity Test                                                                                       21

3.7       Statistical Analysis                                                                                                     22


CHAPTER FOUR

Results                                                                                                                                    22

4.1       Qualitative Phytochemical Screening of Capsicum annuum                                     22

4.2       Quantitative Phytochemical Screening of Capsicum annuum                                    24

4.3       Antimicrobial Activity of Capsicum annuum                                                            26

4.4       Relative Potencies of Capsicum annuum Extract                                                      29


CHAPTER FIVE

Discussion and Conclusion                                                                                                    31

5.1       Discussion                                                                                                                   31

5.2       Conclusion                                                                                                                  33

References

 

 

 

 

 

 

 

LIST OF TABLES

 

Table       Title                                                                Page

4.1                   Qualititative Phytochemical Screening of Capsicum annuum Extract          23

4.2                   Phytochemical Content of Capsicum annuum (%)                                        25

4.3                   Antimicrobial Activities of Capsicum annuum Extract (Diameter

of Zone of Inhibition).                                                                                    27

4.4                   Relative Potency of Capsicum annum extracts antimicrobial activity

Compared with standard antibiotics                                                               30

 

 

 

 

 

LIST OF FIGURES

Figure        List                                                                 Page

1:                     Capsicum annum plant (chili pepper)                                                            6

2:                     Inhibition effect of Capsicum annum extract (Diameter of zone

of inhibition)                                                                                                   28

 


 

CHAPTER ONE

INTRODUCTION


1.1       BACKGROUND OF THE STUDY

There has been an increasing consumer demand for foods free or with low, if any, added synthetic preservatives because synthetic preservatives could be toxic to humans (Bedin, et al., 1999). Concomitantly, consumers have also demanded for wholesome and safe food with long shelf lives. These requirements are often contradictory and have put pressure on the food industry for progressive removal of chemical preservatives and adoption of natural alternatives to obtain its goals concerning safe food with long shelf lives (Brull and Coote, 1999).

Spices and herbs have been used for thousand of the centuries by many cultures to enhance the flavour and aroma of food. Early culture also reorganized the values of using spices and herbs in preventing foods and for their medical values. Spices in the past decade confirm that the growth of both Gram-positive and Gram-negative food-borne bacteria, yeasts and molds can be inhibited by garlic, onion, cinnamon, clove, thyme, sage and other spices. Although, the primary purpose of spices is to impart flavour and piquancy to food, the medicinal, antimicrobial and antioxidant properties of spices have also been exploited (Souza et al., 2005). The antimicrobial activity of spices is documented and interest continues to the present (Uraih, 2004).

The growing concern about food safety has recently led to the development of natural antimicrobials to control food-borne pathogens and spoilage bacteria. Spices are one of the most commonly used natural antimicrobial agents in foods and have been used traditionally for thousands of years by many cultures for preserving foods and as food additives to enhance aroma and flavour (Souza et al., 2005).

Pepper plant is a perennial woody vine growing up to 4 m in height on supporting trees, pole, or trellises. The leaves are alternate, 5 - 10 centimeters long and 3 - 6 cm broad. The flowers are small, produced on pendulous spikes 4 - 8 cm long at the leaf-nodes, the spikes lengthening up to seven to 15 cm as the fruit matures (Azhar et al., 2005). The fruit of the pepper is a drupe and when dried it is a pepper corn. It is grown in soil that is neither too dry nor susceptible to flooding, moist, well-drained and rich in organic matter. The plant is propagated by first planting; it is first planted in the nursery before transplanting the seedling to the farm.

The antimicrobial activities of plants and their extracts against bacteria and fungi have been documented. Plant use in the control management and cure of diseases is an age long practical Mudalige et al., (2011) reported that natural plant products including medicinal plant extracts are being increasingly used as agrochemicals, for the control of diseases. This was attributed to their relative non-phytotoxicity, easy biodegradation and environmental friendliness. Sofoewara, (1993) observed that the use of plant in traditional medications and their effectiveness as antimicrobial agents, stems from their phytochemical composition.

The use of Chilli pepper by humans is an age long practice. Omolo et al. (2014) reported the use of pepper to date back as far as 2500BC. The use of pepper has become an integral part of diet culture of many people, all over the world (Mortensen and Mortensen, 2009).

Botanically, pepper is a member of the plant family of solanaceae, and has close relation to potato, tomato, eggplant, tobacco and petunia (Omolo et al., 2014). It is cultured in the tropics and sub-tropics that are warm and humid. According to the authors, pepper is one of the oldest domesticated crops of the western world and is also currently, the most widely grown spice in the world as it is a major ingredient in most global cuisines.

Although, peppers are used as spice all over the world, their flavour and pungent smell vary widely just as their content of the major active ingredient, capsicum and capsaicinoid analogues vary (Dorantes et al., 2000). According to Seugill, et al. (2014) there is evidence that Chilli peppers were incorporated into a number of medical preparations in mesoamerican regions and the preparations were applied for a variety of ailments including respiratory problems, bowel complaints, ear aches, and sores etc. Again Brito – Argaez et al. (2009) examined the omnipresent nature of pepper in the mesoamerican diets and observed that, it is a spice. Peppers play substantial roles and occupy an important position in their diet culture with very important nutritional consequences. The authors also record the demonstration of high degree of bioactivity by peppers which affects the nervous, cardiovascular and digestive systems, while its chemical composition show high concentration of essential nutrients, including Vitamin C.

In the present world, consumers demand for safe and high quality foods, since there is wide spread information and enlightenment regarding health data and information. The concern for food safety is on the risk due to increasing occurrence of new food – borne disease out breaks caused by pathogenic microorganisms. Also, pathogenic microorganisms are responsible for many other non-food related diseases of humans and live stock and their control with chemical agents has witnessed high level of uneasiness (Cichewicz et al., 1996).

Outside epidermic cases, food-borne pathogens represent a major public health concern (FDA 2013). Again, not much work has been documented on photochemistry and health benefit of local pepper plant, whereas much has been done in the western world on the antimicrobial properties and activities of chilli-pepper plant (mainly on the fruits). It is a reported knowledge that the activity of plant extract against pathogens, stems from their phytochemical composition which in itself  is affected by agronomic factors, since plant synthesize and accumulate there substances with what they are able to absorb from their natural soil environment. Documented records of extract of chilli – peppers and their general and specific antimicrobial activities are many Kalia et al., (2012). Report also show that chilli peppers are receiving a lot of research attention elsewhere, and the capsicum genus is reported to be a good source of antimicrobial and antifungal compounds (Tajkarimi et al., 2010).

Concern has been expressed about the rising prevalence of pathogenic microorganisms which are resistance to the newer modern antibiotics that have been produced in the last three decade (Nascimento et al., 2000). Coincidentally, the last decades has also witnessed increasing intensity studies on extract and biological active compound isolated from plant species used for natural therapies or herbal medicine (Nascimento et al., 2000; Rios and Recio, 2005). For over a thousand year, natural plants have been seen as a valuable source of medicinal agent with proven potentials in the treatment of infectious diseases and with lesser side effects compared to synthetic drug agent (lwu et al., 1999). However the problem pose by high costs, adulteration and increasing side effect of synthetic drug coupled with their inadequacy to diseases treatment especially in the developing countries cannot be ignored (Shariff, 2001).

1.2       OBJECTIVES OF THE STUDY

      i.         To determine the antimicrobial activity of chilli pepper plant leaves against pathogenetic microorganisms.

     ii.         To determine the phytochemical constituents of chilli pepper leaf.


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