TABLE OF CONTENT
Title
page……………………………………………………………………..i
Certification………………………………………………………………….ii
Dedication …………………………………………………………………..iii
Acknowledgment
……………………………………………………………iv
Abstract………………………………………………………………………v
CHAPTER ONE
1.0
Introduction……………………………………………………………..1
1.1
Micro-organisms………………………………………………………....1
1.2 Classification
of micro-organism………………………………………...2
1.2.1
Bacteria………………………………………………………………...3
1.2.2
Virus…………………………………………………………………...4
1.2.3
Algae…………………………………………………………………..4
1.2.4
Fungi…………………………………………………………………..5
1.2.5 Protozoa ………………………………………………………………6
CHAPTER TWO
2.0
Literature review………………………………………………………..7
2.1
Bacteria…………………………………………………………………7
2.2 Bacteria and its cells……………………………………………………9
2.3 Shape of the
bacteria…………………………………………………...12
2.4 Environmental
effect of bacteria……………………………………….14
2.5 Medicinal plants………………………………………………………...16
2.5.1
Characteristics of medicinal plants…………………………………...17
2.6
Medicinal plants as antimicrobial……………………………………...26
2.7 Why
considering medicinal plants as antimicrobial…………………….28
2.8 Botanical profile of Bridelia ferruginea ………………………………30
2.8.1 Taxonomy of plant…………………………………………………...31
2.8.2 Plant description…………………………………………………….31
2.8.3 Properties……………………………………………………………32
2.8.4 Cultivation details…………………………………………………...32
2.8.5 Edible uses…………………………………………………………..32
2.8.6 Medicinal uses……………………………………………………....32
2.8.7 Propagation………………………………………………………….33
2.9 Bridelia ferruginea as
antimicrobial…………………………………….33
CHAPTER THREE
3.0 Materials and
methods………………………………………………….35
3.1 Aims and
objectives…………………………………………………….35
3.2
Materials…………………………………………………………………35
3.2.1
Chemicals,solvents and chemical reagents……………………………35
3.2.2
Equipment and materials………………………………………………36
3.3 Methods………………………………………………………………. 36
3.3.1 Collection
and preparation of plant materials…………………………36
3.3.2 Extraction
of plant materials…………………………………………..36
3.3.3
Phytochemical screening of Methanol extract………………………...37
3.3.4 Phytochemical screening of Hexane
extract………………………….40
3.3.5
Phytochemical screening of Acetone extract…………………………43
3.4 Thin-layer
chromatography……………………………………………47
3.5 Isolation and characterization of the crude
extract……………………49
3.5.1 Column
chromatography………………………………………………49
3.5.2 Mass of
fractions gotten from the column chromatography…………..49
3.6
Instrumentation techniques………………………………………………50
3.6.1 Sample
analysis for GC-MS…………………………………………..50
3.6.2 Sample
analysis for AAS………………………………………………51
3.7 Anti-microbial
activity of Methanol extract of Bridelia ferruginea……………51
3.7.1
Method……………………………………………………………………….51
3.8 Flow diagram
of the entire experiment carried out in this
research
work…………………………………………………………………..52
CHAPTER FOUR
4.0 Result and
discussion ……………………………………………………………53
4.1 Result of
phytochemical analysis of methanol, N-Hexane, and
Acetone extract of Bidelia
ferruginea…………………………………………...53
4.2
Instrumentation techniques……………………………………………………..55
4.2.1 Result for
GC-MS analysis……………………………………………………55
4.2.2 Result for
AAS analysis ………………………………………………………61
4.3 Anti-microbial
activities result…………………………………………………62
CHAPTER FIVE
Conclusion………………………………………………………………………….64
Recommendation…………………………………………………………………..64
Reference
ABSTRACT
Medicinal plants are those plants that are
used in treating and preventing specificailments and diseases that affect human
beings. Hence the important role of medicinal plants in health care
deliverycannot be over emphasized. Brideliaferruginea is atropical medicinal
plant which belongs to the family of euphorbiaceae commonly used in traditional
Africa medicine for treating various diseases. My research in this study was
concentrated on the bark of bridelia ferruginea to detect the active ingredient
responsible for the antimicrobial activities extraction.
Phytochemicalscreening, thin layer chromatography, column chromatography,
GC–MS, AAS and biological activities was carried out on the crude methanol
extract of Bridelia ferruginea. The
pytochemical screening for the methanol crude extract indicate the presence of
tannins, alkaloids, saponins, steroids, resins and phlobatahin, while the
phytochemical screening for acetone crude extract indicate the presence of
tannins, alkaloids, flavonoids, phlobatanin and reducing sugar. And the
photochemical tests on the hexane crude extract gave negative result all
through. Isolation was done using Column chromatography, AAS was used for the
determination of some certain heavy metals present in both the isolate and the
crude which may be toxic to man health and the GC–MS was used to determine the
presences of unknown substances in the sample. Result for GC-MS shown the
presence of seven compounds which are medicinal namely, 2-[4-methyl-6-(2,6,6-trimethylcyclohex-1-enyl)hexa-1,3,5-trienyl]cyclohex-1-en-1-carboxaldehyde,
limonene-6-ol,pivalate, 1-Heptatriacotanol, Benzene,1-4-dicloro, cyclobarbital,
2-nonadecanone 2,4-dinitrophenylhydrazine, 1-monolinoleylglycerol trimethylsilylether.
While for AAS, two metals were detected namely iron (2.1ppm) and zinc (0.05ppm)
which are non-toxic for human consumption. And also result from the
antibacterial activities shows that Bridelia ferruginea was highly susceptible
to Escherichia coli and highly
resistant to Proteus vulgaris.
CHAPTER ONE
1.0 INTRODUCTION
1.1 MICRO-ORGANISM
A
microorganism or microbe is an organism (form of life) that is microscopic (too
small to see by the unaided human eye) [Delong et al 2001]. Is a microscopic
living organism, which may be single-celled or multicellular. The study of
microorganisms is called microbiology, a subject that began with the discovery
of microorganisms in 1674 by Antonie van Leeuwenhoek, using a microscope of his
own design [Madigan M et al 2006].
Microorganisms are
very diverse and include all bacteria, archaea and most protozoa. This group
also contains some species of fungi, algae, and certain microscopic animals,
such as rotifers. Many Macroscopic animals and plants have microscopic juvenile
stages. Some microbiologists also classify viruses as microorganisms, but
others consider these as nonliving. In July 2016, scientists reported
identifying a set of 355 genes from the last universal common ancestor of all
life, including microorganisms, living on Earth [Madigan M et al 2006].
Microorganisms are
often described as single-celled, or unicellular, organisms; however, some
unicellular protists are visible to human eye, and some multicellular species
are microscopic [Delong et al 2001].
Single-celled
microorganisms were the first forms of life to develop on Earth, approximately
3–4 billion years ago. Further evolution was slow, and for about 3 billion
years in the Precambrian eon, all organisms were microscopic. So, for most of
the history of life on Earth, the only forms of life were microorganisms.
Bacteria, algae and fungi have been identified in amber that is 220 million
years old, which shows that the morphology of microorganisms has changed little
since the Triassic period. [Madigan M et al 2006].
Merging the idea of the microscopic and the
very small with the older idea of an organism as a living entity or cell, the
concept of a microorganism enabled a real appreciation of the microbial world
as one that is amenable to study using similar tools and approaches even though
representing distinctly different types of reproductive units and cell
organizations [John A 2014].
1.2 CLASSIFICATION OF MICRO – ORGANISM
Microorganisms can
be found almost anywhere in the taxonomic organization of life on the planet.
Bacteria and archaea are almost always microscopic, while a number of
eukaryotes are also microscopic, including most protists, some fungi, as well
as some animals and plants. Viruses are generally regarded as not living and
therefore not considered as microorganisms, although the field of microbiology
also encompasses the study of viruses [Madigan M et al 2006].
1.2.1 BACTERIA
Bacteria
constitute a large domain of prokaryotic microorganisms. Typically a few micrometers
in length, bacteria have a number of shapes, ranging from spheres to rods and
spirals, bacteria were among the first life forms to appear on earth, and are
present in most of its habitats. Bacteria inhabit soil, water, acidic hot
springs, radioactive waste, and the deep portions of earth’s crust [Rappe M.S
2003].
Most bacteria have
not been characterized, and only about half of the bacterial phyla have species
that can be grown in the laboratory. The study of bacteria is known as
bacteriology, a branch of microbiology. There are approximately ten times as
many bacterial cells in the human flora as there are human cells in the body,
with the largest number of the human flora being in the gut flora, and a large
number on the skin. The vast majority of the bacteria in the body are rendered
harmless by the protective effects of the immune system, and some are
beneficial. However, several species of bacteria are pathogenic and cause
infectious diseases, including cholera,
syphilis, anthrax, leprosy, and bubonic plague. The most common fatal bacterial
diseases are respiratory infections, with tuberculosis alone killing about 2
million people per year, mostly in sub-Saharan Africa [WHO 2002].
In developed countries, antibiotics are used
to treat bacterial infections and are also used in farming, making antibiotic
resistance a growing problem. In industry, bacteria are important in sewage
treatment and the breakdown of oil spills, the production of cheese and yogurt
through fermentation, and the recovery of gold, palladium, copper and other
metals in the mining sector, as well as in biotechnology, and the manufacture
of antibiotics and other chemicals [Ishige T et al 2005].
1.2.2 VIRUS
A virus is a small infectious agent that
replicates only inside the living cells of other organisms. Viruses can infect
all types of life forms, from animals and plants to microorganisms, including bacteria
and archaea. Viruses spread in many ways; viruses in plants are often
transmitted from plant to plant by insects that feed on plant sap, such as
aphids; viruses in animals can be carried by blood-sucking insects. These
disease-bearing organisms are known as vectors. Influenza viruses are spread by
coughing and sneezing. Viruses display a wide diversity of shapes and sizes,
called morphologies. In general,
viruses are much smaller than bacteria.
Most viruses that
have been studied have a diameter between 20 and 300nanometres. Some
filoviruses have a total length of up to 1400 nm; their diameters are only
about 80nm. Most viruses cannot be seen with an optical microscope so scanning
and transmission electron microscopes are used to visualise virions [Breitbart
M et al 2005].
1.2.3 ALGAE
Algae is an informal
term for a large, diverse group of photosynthetic organisms which are not
necessarily closely related, and isthus polyphyletic. Included organisms range from
unicellular genera, such as Chlorella and
the diatoms, to
multicellular
forms, such as the giant kelp, a large brown alga which may grow up to 50 m in
length. Most areaquatic and autotrophic and lack many of the distinct cell and
tissue types, such as stomata, xylem, and phloem, which are found in land
plants. The largest and most complex marine algae are called seaweeds, while
the most complex freshwater forms are the Charophyta, a division of green algae
which includes, for example, Spirogyra and
the stonewort’s. Most algae contain chloroplasts that are similar in structure
to cyanobacteria. Chloroplasts contain circular DNA like that in cyanobacteria
and presumably represent reduced endosymbiotic cyanobacteria [Allaby M. 1992].
1.2.4 FUNGI
Fungi is any
member of a group of eukaryotic organisms that includes organisms that includes
microorganisms such as yeasts and molds, as well as the more familiar mushrooms.
These organisms are classified as kingdom fungi, which is separate from the
other eukaryotic life kingdoms of plants and animals.
A characteristic
that places fungi in a different kingdom from plants, bacteria, and some
protists is chitin in their cell wall. Similar to animals, fungi are
heterotrophs; they acquire their food by absorbing dissolved molecules,
typically by secreting digestive enzymes into their environment [Wikipedia
2016].
The fungi have
several unicellular species, such as baker's yeast and fission yeast. Some
fungi, such as the pathogenic yeast, can undergo phenotypic switching and grow
as single cells in some environments, and filamentous hyphae in others [Madigan
M et al 2006].
Along with
bacteria, fungi are responsible for the decomposition and reprocessing of vast
amounts of complex organic matter; some of this is recycled to the atmosphere
as CO2, while much is rendered into a
form that can be utilised by other organisms [john wiley et al 2005].
1.2.5 PROTOZOA
Protozoa, as traditionally defined, are mainly
microscopic organisms, ranging in size from 10 to 52 micrometers. Some,
however, are significantly larger. Among the largest are the deep-sea–dwelling
xenophyophores, single-celled foraminifera whose shells can reach 20 cm in
diameter. Free-living forms are restricted to moist environments, such as
soils, mosses and aquatic habitats, although many form resting cysts which
enable them to survive drying. Many protozoan species are symbionts, some are
parasites, and some are predators of bacteria, algae and other protists
protozoawere defined as single-celled animals or organisms with animal-like
behaviors, such as motility and predation. The group was regarded as the
zoological counterpart to the "protophyta", which were considered to
be plant-like, asthey are capable of photosynthesis. The terms protozoa and protozoans are now mostly used informally to designate
single-celled, non-photosynthetic protists, such as the ciliates, amoebae and
flagellates. The term Protozoa was introduced in 1818 for a taxonomic class,
but in later classification schemes the group was elevated to higher ranks,
including phylum, subkingdom and kingdom [Protozoa 2006].
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