ANTIMICROBIAL ACTIVITY AND PROXIMATE COMPOSITION OF SOME SPICES

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ABSTRACT

The present study was conducted to evaluate the anti-microbial and proximate composition of sixteen different spices. The spices investigated were Monodora myristicaCoranarum sativumLaurus nobilisCurcuma longaPiper guineeseRosmarinus officinalisFoeniculum villgareParinari excelsaChrysobalanus icacoZingiberofficinaleTrigonella foenum-gruecumCumin cyminumCarum carviRianodendum heudelotiiAfrostyrax lepidophylus and Afromomum subsericeum. The microorganisms used for the anti-microbial assay were seven clinical pathogens; Escherichia coliStaphylococcus aureusBacillus subtilisProteus vulgarisKlebsiella pneumoniaPseudomonas aeruginosa and Enterococcus faecalis; two of these were used for disc diffusion assay and seven for the minimum inhibitory concentration. Proximate analysis result revealed the moisture content of the samples ranging between 10.24 – 24.67, dry matter 88.30 – 89.76, ash content 3.54 – 9.43, crude protein 7.52 – 17 18, fat 2.67 – 7.95, crude fibre 6.46 – 9.52, and carbohydrate 123.29 – 137.62. The antimicrobial screening revealed that all the spices used in this study have good antibiotic properties against the various bacteria pathogens tested with maximum inhibition observed in Parinari excelsa (24mm) and Cumin cyminum (24mm) in the disc diffusion assay and Curcuma longa (7.18uglmlin the MIC. The result of this study showed that all the selected spices contained reasonable amount of nutraceutical values which suggest the application of the seeds as supplementary sources of antimicrobial agents and essential nutrients.







TABLE OF CONTENTS

Title page                                                                                                                                i

Declaration                                                                                                                             ii

Certification                                                                                                                           iii

Dedication                                                                                                                              iv

Acknowledgements                                                                                                                v

Table of contents                                                                                                                    vi

List of tables                                                                                                                           xi

List of plates                                                                                                                           xii

Abstract                                                                                                                                  xiii

CHAPTER ONE: INRODUCTION

1.1       Background of the study                                                                                             1

1.2       Objectives of study                                                                                                     5

1.3       Justification of the study                                                                                             6

CHAPTER TWO: LITERATURE REVIEW

2.1       Spices                                                                                                                          7

2.2       History of spices                                                                                                         7

2.3       Conventional classification of spices                                                                         7

2.4       Importance of spices                                                                                                   8

2.5       Potential nutraceutical ingredients from plant origin                                                 10

2.5.1    Guinea pepper (Piper guineense)                                                                               10

2.5.1.1 Origin, habitat and distribution of P. guineense                                                         10

2.5.1.2 Botany of the plant                                                                                                     11

2.5.1.3 Chemical composition                                                                                                11

2.5.1.4 Nutraceutical values of Piper guineense                                                                    13

2.5.2    African nutmeg (Monodora myristica)                                                                      14

2.5.2.1 Origin and Distribution                                                                                              14

2.5.2.2 Botany of the plant                                                                                                     14

2.5.2.3 Chemical composition                                                                                                15

2.5.2.4 Nutraceutical values of M. myristca                                                                           15

2.5.3    Ricinodendron heudelotii (Njasang)                                                                          17

2.5.3.1 Botany of the Plant                                                                                                     17

2.5.3.2 Nutraceutical values of Ricinodendron heudelotii                                                     18

2.5.4    Country onion (Afrostyrax lepidophyllus)                                                                 18

2.5.4.1 Chemical composition of the plant                                                                             19

2.5.4.2 Nutraceutical values of Afrostyrax lepidophyllus                                                      19

2.5.5    Coriandrum sativum Linn. (Coriander)                                                                     20

2.5.5.1 Origin and distribution of the plant                                                                            20       

2.5.5.2 Botany of the plant                                                                                                     21

2.5.5.3 Chemical constitutes of Coriander                                                                             21

2.5.5.4 Nutraceutical values of Coriandrum sativum                                                             23

2.5.6    Fenugreek (Trigonella foenum−graecum L.)                                                             23

2.5.6.1 Botany of the Plant                                                                                                     25

2.5.6.2 Chemical compositions of Fenugreek                                                                        25

2.5.6.3 Nutraceutical values of Fenugreek                                                                             25

2.5.7    Cumin (Cuminum cyminum)                                                                                      26

2.5.7.1 Origin and Distribution                                                                                              27

2.5.7.2 Botany and Description of the Plant                                                                          27

2.5.7.3 Chemical constituents                                                                                                 28

2.5.7.4 Nutraceutical values of Cumin                                                                                   29

2.5.8    Ginger (Zingiber officinalis)                                                                                     29

2.5.8.1 Origin of the Plant                                                                                                      30

2.5.8.2 Botany of the Plant                                                                                                     30

2.5.8.3 Chemical Constituents of Ginger                                                                               30

2.5.8.4 Nutraceutical values of Ginger                                                                                   31

2.5.9    Tumeric (Curcuma longa L.)                                                                                      32

2.5.9.1 Botany of the Plant                                                                                                     32

2.5.9.2 Chemical Constituents and Therapeutic Potentials of Turmeric                                    33

2.5.9.3 Nutraceutical values of Curcuma longa                                                                     33

2.5.10  Bayleaf (Laurus nobilis L.)                                                                                        34

2.5.10.1 Origin and Distribution                                                                                            35

2.5.10.2 Botany of the Plant                                                                                                   35

2.5.10.3 Chemical Composition                                                                                             36

2.5.10.4 Nutraceutical values of L. nobils                                                                              36

2.5.11  Rosmarinus officinalis (Rosemary)                                                                            37

2.5.11.1 Chemical Composition                                                                                             37

2.5.11.2 Nutraceutical values of R. officinalis                                                                       38

2.5.12  Parinari excelsa                                                                                                         39

2.5.12.1  Nutraceutical values of Parinari excelsa                                                                40

2.5.13  Chrysobalanus icaco (Coco plum)                                                                             40

2.5.13.1 Botany of the Plant                                                                                                   41

2.5.13.2 Nutraceutical values of Chrysobalnus icaco                                                                        41

2.5.14  Caraway (Carum carvi)                                                                                              42

2.5.14.1 Origin and Distribution                                                                                            42

2.5.14.2 Botany of the Plant                                                                                                   42

2.5.14.3 Chemical Composition                                                                                             43

2.5.14.4 Nutraceutical values of C. carvi                                                                               43

2.5.15  Fennel (Foeniculum vulgare Mill.)                                                                            44

2.5.15.1 Origin and Distribution                                                                                            45

2.5.15.2 Botany of the Plant                                                                                                   45

2.5.15.3 Chemical Composition of Fennel                                                                             46

2.5.15.4 Nutraceutical Value of Foeniculum vulgare                                                            47

2.5.16  Afromomum subsericeum (Ataiko)                                                                             48

2.5.16.1 Origin and distribution                                                                                             48

2.5.16.2 Botany                                                                                                                      48

2.5.16.3 Nutraceutical values of the plant                                                                              48

2.6       Antimicrobial susceptibility testing                                                                           49

2.7       Antimicrobial assays                                                                                                  49

2.8       Proximate analysis                                                                                                      50

CHAPTER THREE: MATERIALS AND METHODS   

3.1       Sample collection and identification                                                                          52

3.1.1    Preparation of extracts                                                                                                52

3.1.2    Collection of microorganisms                                                                                    52

3.2       Evaluation of spices for antimicrobial activity                                                          53

3.2.1    Preparation of stock/working solution                                                                       53

3.2.2    Preparation of test organism for susceptibility                                                          53

3.2.3    Media preparation                                                                                                       53

3.2.4    Determination of antimicrobial activity of extracts (Disc Diffusion Method)          54

3.2.5    Determination of minimum inhibitory concentration (MIC)                                     54

3.3       Proximate analysis                                                                                                      54

3.3.1    Determination of moisture content                                                                             54

3.3.2    Determination of crude fat                                                                                         55

3.3.3    Determination of crude fiber content                                                                         55

3.3.4    Determination of crude protein content                                                                     56

3.3.5    Determination of ash content                                                                                     56

3.3.6    Determination of carbohydrate content                                                                      57

 

 

CHAPTER FOUR: RESULTS

4.1       Description and biological activities of plants used as spices obtained during

survey                                                                                                                         58

4.2       Antimicrobial activity of methanol spice extract of selected spices on two

bacterial isolates                                                                                                         60

4.3       Minimum inhibitory concentration (MIC) of methanolic spices extract against

seven (7) bacterial pathogen                                                                                       63

CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATIONS

5.1       Discussion                                                                                                                   68

5.2       Conclusion                                                                                                                  68

5.3       Recommendations                                                                                                      68

References







 

LIST OF TABLES

Table                                                  Title                                            Page

1                Conventional classification of spices                                                             8

2                Ethnobotanical Survey of Plants used as spices in South-East Nigeria                  58

3                Antimicrobial activity of methanolic extracts of selected spices on two

isolated microbes represented by disc diffusion method in (mm)                     61

4                MIC result of methanolic extracts of the plant samples against seven

isolated pathogens and are represented in ug/ml                                            65

5                Proximate Composition of Spices (%)                                                           67





LIST OF PLATES

Plate                                                   Title                                              Page

1                      Picture of Guinea pepper (Piper guineense)                                                  10

2                      Picture of Monodora myristica                                                                       14

3                      Picture of Ricinodendron heudelotti                                                              17

4                      Picture of Afrostyrax lepidophyllus                                                                18

5                      Picture of Coriandrum sativum                                                                      20

6                      Picture of Trigonella foenum−graecum                                                         23

7                      Picture of Cuminum cyminum                                                                        26

8                      Picture of Zingiber officinalis                                                                        29

9                      Picture of Curcuma longa                                                                              32

10                    Picture of Laurus nobilis                                                                                34

11                    Picture of Rosmarinus officinalis                                                                   37

12                    Picture of Parinari excels                                                                               39

13                    Picture of Chrysobalanus icaco                                                                     40

14                    Picture of Carum carvi                                                                                   41

15                    Picture of Foeniculum vulgare                                                                       44

16                    Picture of Afromomum subsericeum                                                               48

17                    Escherichia coli                                                                                        62

18                    Staphylococcus aureus                                                                                   62

19                    Pictures of Microtitre plates showing MIC results of the plant samples

against seven (7) microorganisms viz: Enterococcus, Staphylococcus,

Bacillus, Klebsiela, Proteus, Pseudomonas and E. coli respectively     66

 


 



 

CHAPTER ONE

1.0       INTRODUCTION

1.1       BACKGROUND OF THE STUDY

Spices are defined by Corn (1999) as dried seeds, fruits, roots, barks, leaves or vegetables used nutritionally in a very small quantity as food additives for the purpose of flavor, color or as preservative that kills harmful bacteria or suppress their growth. Spices are also products from plants seeds, fruits, flowers, and roots or bark that are added to food to improve flavor, taste, and color or act to minimize the rate of rancidity and as preservatives that suppress microbial activities (Thomas, 2007). In Nigeria, a high proportion of the rural and urban population resort to natural food ingredients particularly because of their availability. Spices are a large group of such ingredients. Some spices are used for the preparation of special types of soup, these include: soup for newly delivered mothers to accelerate blood flow leading to the elimination of blood cloths from her womb and blood system. Some have been recommended for fast relief of ailment such as cholera, diarrhea, dysentery and wound sepsis (Inyang, 2003, OlumSimbo et al., 2011).  They are also used as vegetables for spicing meat, oil bean salad and foods. Most of these spices have been associated with abundant bitter principle which is claimed to reduce blood sugar levels and the liquor takes as a purge for colic, stomach pains and worm infections (Friday et al., 2011).

Each spice has a unique aroma, flavor and antimicrobial activities which are derived from phytochemicals and some spices are reported to have microbiocidal or microbiostatic activities (Onwuliri and Wonang, 2005). More recently the interest in spices has grown not only for the seasoning and flavouring properties but also the antioxidant potential (Mukul et al., 2013). Spices are rich in phytochemicals and other active ingredients that protect against disease and promote healing (Pavitra, 2013). It is also believed that new born babies grow rapidly when they are fed with food made of these spices (Roger, 2002).

Infectious diseases caused by pathogens and food poisoning caused by spoilage microorganisms are threatening human health all over the world. The efficacies of some antimicrobial agents which are currently used to extend shelf life and increase the safety of food products in food industry and to inhibit disease causing microorganisms in medicine have been weakened by microbial resistance (Qing et al., 2014). Also food borne illness caused by consumption of foods contaminated with pathogenic bacteria or the toxins has been of great public health concern. In recent times, consumers are even more concerned of the processed foods they eat not only because of the high risk of contamination but also because of the added synthetic preservatives which may be hazardous to health. Food additives such as monosodium glutamate, aspartame, saccharine, sodium cyclamate, sulfites, nitrates, nitrites and antibiotics have all been reported to cause clinical conditions manifesting variously as headache, nausea, weakness, mental retardation, seizures, cancer and anorexia (Rangan and Barceloux, 2009; Wroblewsca, 2009). In recent years the prevention of cancer and cardiovascular diseases has been associated with the ingestion of spices, fresh fruits, vegetables or teas rich in natural antioxidnats (Virgili et al., 2001). The protective effects of plant products are due to the presence of several components which have distinct mechanisms of action; some are enzymes and proteins and others are low molecular weight compounds such as flavonoids (Zhang and Wang, 2002). The story of spices and other flavouring materials is one of the most interesting chapters in the history of vegetable products (Obadoni and Ochuko, 2002). The cravens for spices have been one the great factors in human progress and have done much to change the course of history and geography and to promote international relations (Akindahumsi and Salawu, 2005). These spices are said to be therapeutically useful in the management of convulsion, leprosy, stomach ache, inflammation and rheumatoid pains, cough and loss of appetite (Valko et al., 2007).   

The proximate and nutrient analysis of medicinal plants, edible fruits and vegetables plays a crucial role in accessing their nutritional significance (Pandey et al., 2006). As various medicinal plants species are consumed as food along with their medicinal benefits, evaluating the nutritional significance can help to understand the worth of these plant species (Pandey et al., 2006). There are also various claims about the usefulness of some of these spices especially their use in fattening homes and remarkable growth of new born babies whose mothers use these spices. 

Proximate analysis of plants plays a crucial role in assessing their nutritional significance. Carbohydrates, fats, fibre and protein are the essential nutrients of life. The quality and quantity of proteins in plants are basic factors and important for the selection of plants for nutritive value, systematic classification and plant improvement programs (Nisara et al., 2009). As various spices are used in food along with their medicinal benefit, evaluating the nutritional significance can help understand the worth of these plant species. 

The statement “let food be thy medicine, thy medicine shall be thy food”, made by Hippocrates, the father of modern medicine, is receiving a lot of interest today as food scientists and consumers realize the health benefits of certain foods (El Sohaimy, 2012). The concept of functional foods was invented in Japan. In the 1980s, health authorities in Japan recognized the need for an improved quality of life as well as increasing life expectancy for the number of elderly people in population in order to control health care cost.

According to European Food Information Council, functional foods must contain biologically active components that have the potential to optimize physical and mental well-being and which may also reduce the risk of disease

The term “nutraceutical” was coined from the words “nutrition” and “pharmaceutical” in 1989 by Stephen De Felice, founder and chairman of the foundation for innovation in Medicine (FIM), Granford. He defined nutraceuticals as a food or part of a food that provides medicinal or health care benefits, including the prevention and for treatment of a disease.

Nutraceuticals can be organized in several ways depending upon its easier understanding and application. Some of the most common ways of classifying nutraceuticals can be on food sources, mechanism of action, chemical nature etc. the food sources used nutraceuticals are all natural and can be categorized as (Kalia, 2005; Kokate et al., 2002):

1.     Dietary fiber

2.     Probiotics

3.     Prebiotics

4.     Polyunsaturated fatty acid

5.     Antioxidant vitamins

6.     Polyphenols

7.     Spices

Spices are esoteric food adjuncts that are used for thousands of years to enhance the sensory quality of food (Lipi, 2012). The quantity and the variety of the spices consumed in the tropical countries are particularly extensive. These impart characteristic flavor, aroma, or piquancy and color to foods, stimulating our appetite as well as modify the texture of food. Recent research reveals that dietary spices in their minute quantities has immense influence on the human health by their antioxidative, chemopreventive, anti-mutagenic, anti-inflammatory, immune modulatory effects on cells and a wide range of beneficial effects on human health by the action of gastrointestinal cardiovascular, respiratory, metabolic, reproductive, neural and other systems (Kochhar, 2008).

Most of the spices components are terpenes and other constituents of essential oils. They have been found to be effective in different forms. For instance, Fenugreek seeds (25-50 g), garlic (5-6 cloves), onion (50 g) and turmeric powder (1 pinch) in the daily diet of diabetics prevent and manage long-term complications of diabetes. Regular intake of curcuminoids at about 0.5 g reduces blood lipid peroxide level up to about 3% due to their antioxidant activity (Sreejayan and Rao, 1994). Spices and herbs are in most cases harmless, when used as food, but may exhibit toxicity, when used as medicine, because higher dose administered, or rather due to the possibilities of their interactions with other pharmaceutical medications (Ernst, 2003).


1.2       OBJECTIVES OF STUDY

The specific objectives of the study are;

1.     To assess the various spices and their usage especially the local spices.

2.     To determine the medicinal values−antimicrobial properties of the spices. A mini survey is to be conducted among spices sellers in three selected markets in Abia State.

3.     To determine their nutritional value−proximate properties of the spices.


1.3       JUSTIFICATION OF THE STUDY

Malnutrition and microbial outbreak are major challenges to human health both in rural and urban centre, most especially in developing country the Nigeria. Bacterial and fungal pathogens are the main cause of severe infection in humans and microbes’ resistance to antibiotic has become a serious problem of public health. According to world health organization (WHO), eighty percent of world’s population used medicinal plant as the main primary health care source in the treatment of several diseases. In African countries the rate is much higher (Ajose, 2007). 

Limited scientific evidence regarding safety and efficacy to support the continued therapeutic application of some of these herbal remedies exist compared to such evidence for synthetically formulated drugs (Sowemimo et al., 2007). With the upsurge in the use of herbal remedies, there is need for thorough scientific evaluation to validate or disprove the supposedly therapeutic effect of some of these medicinal plants. However, some of the spices used in this study have not been thoroughly investigated for antimicrobial efficacy and proximate composition. Hence there is need to study the antimicrobial and proximate compositions of these spices. The current is therefore aimed at investigating in nutritional value and antimicrobial activities of some the spices used.

 

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