ABSTRACT
The antimicrobial activity of aqueous and ethanolic extract of leaf and seed of piper guineense was determined on three bacteria (Staphylococcus aureus, Escherichia coli and Salmonella spp) and three fungi (Aspergillus niger, Aspergillus flavus, and Pencillium spp) using agar well diffusion method and minimum inhibitory concentration (MIC). The ethanolic extract was found to show more activity than the aqueous extract on the isolates. The diameter zone of inhibition for the ethanolic extract (leaf and seed) range between 8 and 21mm while that of aqueous extract ranged between 8 and 17mm. The MIC of the ethanolic extract ranged from 3.125 to 100mg/ml, while for aqueous extract, the MIC was from 3.125 to 25mg/ml. Escherichia coli was found to show the greatest sensitivity of all the isolates while Penicillium spp showed the least sensitivity of all the isolates. This observed difference showed that Piper guineense extracts contains antibacterial and antifungal compounds that may be useful for the discovery of new antibiotics. The phytochemical analysis carried out on the Piper guineense leaf and seed revealed the presence of alkaloids, tannis, saponins, glycosides and flavonoids. The presence of these phytochemicals supports the use of this plants as an antimicrobial agent.
TABLE
OF CONTENTS
Title Page i
Certification ii
Dedication iii
Acknowledgements iv
Table of Contents v
List of Tables vii
Abstract viii
CHAPTER ONE
1.0
INTRODUCTION 1
1.1 Aims and Objectives 3
CHAPTER TWO
2.0 LITERATURE REVIEW 4
2.1 Morphology and Description of Piper
guineese 5
2.2 Nutritional value of P.
guineense 6
2.3 Non medicinal Uses of Piper Guineense 6
2.4 Ethnomedicinal Uses of Piper
Guineense 7
2.5 Antimicrobial Activity of Piper
Guineense 8
2.6 Major groups of Antimicrobial
Phytochemicals from Plants 10
2.7 Phytochemical of Piper
Gineense 13
2.8 Description of Test Organism 14
CHAPTER THREE
3.0 MATERIALS AND METHODS 19
3.1 Collection and Identification of Plant Material 19
3.2 Sterility 19
3.3 Test Organism 19
3.4 Identification of Bacteria 19
3.4.1 Gram reaction 19
3.4.2 Biochemical Test 20
3.5 Identification of isolated fungi 22
3.6 Sample Preparation 23
3.7 Extraction Procedures 23
3.8 Media Preparation 23
3.8.1 Preparation of Crude Plant Extracts 23
3.8.2 Determination of Antimicrobial Activity of Extracts 24
3.8.3 Determination of Minimum Inhibitory
Concentration (MIC) of Crude Extracts 24
3.8.4 Phytochemical Screening of the Plant
Extract (leaf and seed) 25
CHAPTER FOUR
4.0
RESULTS 27
CHAPTER FIVE
5.0 Discussion, Conclusion, and Recommendation 51
References 55
LIST OF TABLES
TITLE
PAGE
Table 1: Biochemical Identification of the Test
Organisms 28
Table 2: Cultural
Morphology and Microscopic Characteristics Test Fungi 30
Table 3: Antimicrobial activity of the Piper guineense (Uziza) leaf ethanol extract against test organisms 32
Table 4: Antimicrobial activity of the Piper guneense (Uziza) leaf water
4 extract against test organisms 35
Table 5: Antimicrobial activity of the Piper guneense (Uziza) seed ethanol extract against test organisms 36
Table 6: Antimicrobial activity of the Piper guneense (Uziza) seed water extract against test organisms 39
Table 7: Zone of inhibition (mm) of standard antimicrobial agent (antibiotics) against the test
organism’s positive control (Streptomicin and Nystanin) 40
Table 8: Minimum inhibitory concentration of Piper guneense (Uziza) seed extract against test bacteria 42
Table 9: Minimum inhibitory concentration of Piper guneense (Uziza) leaf extract against test bacteria 44
Table 10: Minimum inhibitory concentration of Piper guneense (Uziza) seed extract against test fungi 46
Table 11: Minimum inhibitory concentration of Piper guneense (Uziza) leaf extract against test fungi 48
Table 12: Qualitative Phyto-Chemical Constituents of Piper guneense (Uziza) Plant Extract 50
CHAPTER ONE
1.0 INTRODUCTION
Food borne illness caused by consumption of
foods contaminated with pathogenic bacteria or their 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, saccharin,
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; Wroblewska, 2009). The increasing demand for food with longer
shelf life, food with little or no chemical preservatives coupled with the
concern about toxic effects of some preservatives has resulted in increased
pressure to find alternatives for better healthcare. Therefore, there is a considerable interest
to stop the disease outbreaks caused by pathogenic and/or spoilage food
microorganisms among food processors, food safety researchers and regulatory
agencies (Marij et al., 2009).
An antimicrobial is any
substance of natural, semisynthetic or synthetic origin that kills or inhibits
the growth of microorganisms but causes little or no damage to the host. Antimicrobial
agents, are a large variety of chemical compounds and physical agents
that are used to destroy microorganisms or to prevent their development (Lai et al., 2004). Antimicrobial agents of
plant origin have been documented and spices are among those perceived to have
great potentials for use as antimicrobial agents (Arora and Kaur, 1999; Okeke et al., 2001). Spices are defined by Corn (1999) as dried
seeds, fruits, roots, barks, leaves or vegetables used in nutritionally
insignificant quantities as food additives for the purpose of flavour, colour
or as preservative that kill harmful bacteria or suppress their growth.
Spices, which include plant materials of
medicinal importance, have been used for the treatment of human ailments way
back in the history of man. In Nigeria, 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 clots from
her womb and blood system. Some have been recommended for fast relief of
ailments such as cholera, diarrhoea, dysentery and wound sepsis (Inyang, 2003,
Olumsimbo et al., 2011).
It is now recognized that spices and herbs
may fulfil more than one function in foods to which they are added. These
include imparting flavour, prolonging the storage life of foods by their
bacterostatic or bacterocidal activity, in addition to being nutrients. These
appeal to consumers who tend to question the safety of synthetic food additives
(Eruteya and Odunfa, 2009). The medicinal and preservative values of spices
have been attributed to the presence of bioactive antimicrobial compounds (Lai
and Roy, 2004).
Piper
guineense (Igbo: Uziza) is a
flowering vine in the family Piperaceae, cultivated for its fruit which is
usually dried and used as spice for seasoning. In the dried form the fruit is
often referred to as peppercorn or simply pepper. Pepper gets its spicy heat
mostly from the piperine compound which has been reported to exhibit
antimicrobial properties detectable both in the outer fruit and in the seed
(Oladosun et al., 2012).
The fruits of the guinea pepper (Piper guineense, Uziza in Igbo) is a
common spices and condiments included in a variety of indigenous Nigerian
recipes particularly among the Igbos (Okeke et
al., 2001). In a recent survey, respondents in the region indicated that
the spice act as a stimulants and laxatives, used to smoothen the skin and cure
fever, cough and stomach disorders. They are also used as abortificients to
treat amenoria and cleanse the womb after childbirth (Okeke, 1998).
1.1 AIMS AND OBJECTIVES
The aim
of this research is to determine the antimicrobial effect of piper guineense (uziza against some food
contaminating organisms and to determine the quantity of the spices needed to
inhibit the growth of the organisms.
The
Objectives are;
To determine the antimicrobial activity of
the spices against a variety of test organisms
To determine the minimal concentrations of
the spices needed to inhibit the growth of the organisms.
To determine the phytochemical compounds
present in the leave and seed of the piper
guineense (uziza).
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