ABSTRACT
Investigations were carried out on mellisopalynological analysis of honey produced by West African honeybee (Apis mellifera, Adansonii latereille) in selected guinea savanna states of Nigeria. Fresh honey samples from two sites (Nassarawa state and Plateau state) in guinea savanna zone of Nigeria were analyzed for pollen spectra, physico-chemical parameters, composition (abundance of pollen), phyto-chemical composition and microbial activities through the zone of inhibition (ZOI). The result showed that pollen grains of myrtaceae and syzygium recorded the most abundant of 40.06 pollen grains in honey samples collected from Nassarawa state and 16.55 in Plateau state. 12 pollen grains of different plant species were identified in Nassarawa which are P. biglobosa, Dicranolepsis usambarica, Ceiba pentandra, Bombax buonopozense, Hygrophylia spp,Grewia spp, Azadirachta indica, Combretaceae/melastomataceae, Vernonia spp, Parkia bicolor, Ludwigia repens and Peltophorum pterocarpum. Ten pollen grains and two fungal spores were identified which are Syzygium guiniense, Eucalyptus spp, Asteraceae, Khaya senegalensis, Lactuca spp, Poaceae, Khaya spp, Diospyros spp, Mangifera indica, Sida acuta, Fungal spore and Sporaschisma spp were identified in Plateau while eight pollen grains of different families identified in the two study sites which includes Syzygium guiniense, Eucalyptus spp, Asteraceae, Parkia biglobosa, Dicranolepsis usambarica, Ceiba pentandra, P. biglobosa and Acanthaceae. Among the examined physico-chemical parameters, viscosity showed the highest mean value of 18.43 in Nassarawa state and 16.02 in Plateau state and Density with the lowest mean value of 1.41 in Nassarawa state and 1.41 in Plateau state. Colour intensity of 417.69 and 408.93 were recorded in the two study sites which indicated that the honey samples are dark amber in colour according to USDA, (1985). The moisture content of Honey sample ranged between 17.80-19.85 and 19.95-19.85 in the two states. The pH range (3.92-4.15 and 3.07-3.88) obtained in this study showed that all honey samples from the two states were acidic. Ash content of 0.28-0.34 for Nassarawa state and 0.34-0.37 for Plateau State was recorded. Crude protein of 0.24-0.32 and 0.21-0.37 was also recorded in the study sites. The macro nutrients observed includes Calcium, Nitrogen, Phosphorus, magnesium and Potassium where potassium had the highest value of 54.92 and 58.90 while nitrogen had the least mean value of 0.064 and 0.074 in the two states. The metals in the honey samples were present in small amounts. The range percentage Glucose, Fructose, Sucrose and TSS from the study was 21.39-25.28%, 32.61-2.79%, 2.32-2.79% and 62.09-63.43% for Nassarawa and 19.17-23.29% 28.88-31.39%, 2.30-2.79% and 62.09-65.81% for Plateau state. The result revealed that both honey samples contained vitamin B1, B2, B3, E, and B9. The result also revealed that all the bacteria and fungi species (Pseudomonas aeruginos,Escherichea coli, and Salmonella spp.) and (Rhizopus, Penecillium citrinum and Aspergillus niger) investigated were sensitive to the honey samples at all concentration levels and the positive control gave the highest zone of inhibition.
TABLES OF CONTENTS
Title page i
Declaration ii
Certification iii
Dedication iv
Acknowledgements v
Table of content vi
List of figures x
List of plates xi
List of tables’ xii
Appendices
xiii
Abstract xiv
CHAPTER 1: INTRODUCTION
1.1 Statement of Problems 3
1.2 Aims and Objective 3
1.3 Justification 3
1.4 Scope and Limitation of Study 4
CHAPTER 2: LITERATURE REVIEW
2.1 Plant Identification 5
2.1.1 Pollen 5
2.1.2. Pollen analyses of honey 5
2.2 Honey 6
2.2.1 Honey
bees 6
2.2.2 Categories
of honeybee 7
2.2.3 Beekeeping 8
2.2.4 Beeswax 9
2.2.5 Propolis
and Royal jelly 9
2.3 Honey
Composition and Its Uses 10
2.3.1 Honey
composition 10
2.4 Physical
and Chemical Characteristics of Honey 11
2.4.1 Viscosity 11
2.4.2 Density 11
2.4.3 Hygroscopicity 11
2.4.4 Color 12
2.4.5 Crystallization 12
2.4.6 Hydroxyl-methyl-furfural
(HMF) 13
2.4.7. PH
and free acidity 13
2.5 Analyses
of Modern and Traditional Bee Keeping 14
2.6 Contaminants
from Environmental Sources 15
2.6.1 Heavy
metals 15
2.6.2 Organic contaminants 15
2.6.3 Other
contaminants 16
2.6.4
2.6.4 Conventional Knowledge of honeybee keeping (Honey Hunting) 16
2.6.5 Modern beekeeping 17
2.7 Hive type Need not Determine Honey
Quality 17
2.7.1 Fixed comb hives 18
2.7.2 Movable
comb hives 18
2.7.3 Movable-frame
hives 18
2.8 Harvesting
of honey 19
2.9 Role of Beekeeping in Forest
Sustainability 19
2.10 Benefits of Beekeeping in
Livelihood Activity 21
2.10.1 Enhancing local skills, knowledge and
traditions 21
2.10.2 Integration into the farming system 21
2.10.3 Dietary
contribution 22
2.10.4 Traditional
medicinal value 22
2.10.5 Beekeeping
and farm processing 23
2.10.6
Improved income of beekeeping 24
2.10.7 Social
benefits of beekeeping 24
2.10.8 Environmental
benefits of beekeeping 25
CHAPTER 3: MATERIALS AND METHOD
3.1 Study
Area 26
3.2 Materials 31
3.3 Methodology 31
3.3.1 Inventory
and field survey 34
3.3.2 Procedure
for honey sample collection 34
3.3.2.1 Identification
of plants visited by honey bees 35
3.4 Pollen Analysis 36
3.4.1 Extraction of Pollen from Samples of Honey 36
3.4.2 Acetolysis 36
3.4.3 Temporary Slides 37
3.4.4 Pollen identification and composition 37
3.5 Proximate Analysis 37
3.5.1 Determination of moisture content 38
3.5.2 Determination of total ash 38
3.5.3 Determination of Crude Fibre 39
3.5.4 Determination of protein 39
3.5.5 Determination of fat content 40
3.5.6 Determination of carbohydrates 41
3.6 Determination of Minerals 41
3.6.1 Determination of Phosphorus 41
3.6.2 Determination
of Calcium and Magnesium 42
3.6.3 Determination
of Potassium and Sodium 43
3.7 Heavy
Metal Determination 44
3.8 Determination
of Vitamins in Honey Sample 44
3.8.1 Determination
of Folic Acid (B9) 44
3.8.2 Determination
of Riboflavin (B2) 45
3.8.3 Determination of Thiamine 45
3.8.4 Determination of Niacin 46
3.9 Quantitative Determination of The
Phytochemical Constituents Of Honey 47
3.9.1 Alkaloid
Determination 47
3.9.2 Flavonoid
Determination 47
3.9.3 Phenols
Determination 48
3.9.4 Saponins
Determination 49
3.9.5 Steroid
Determination 49
3.9.6 Tannin
Determination 50
3.10 Microbial
Analysis 51
3.10.1 Preparation
of microorganism for the experiment 51
3.10.2 Sample
Preparation 51
3.10.3 Sensitivity
Test of honey on the microorganism 51
CHAPTER 4: RESULT AND
DISCUSSION
4.1 The Composition and Abundance of Pollen Grains
Identified in Honey Samples 53
4.2 Pollen Spectra In Honey Sample
From Study Areas 53
4.3 Physicochemical Compositions Of
Honey Samples From Study Area 54
4.3.1Physical
composition of honey samples 58
4.3.2 Macro
and micro nutrient of honey samples 63
4.3.3 Percentage
sugar content in honey samples 66
4.3.4 Phytochemical
composition of honey samples 67
4.3.5 Vitamin
composition of honey samples 67
CHAPTER 5: CONCLUSION AND RECOMMENDATIONS
5.1 Conclusion
78
5.
2 Recommendation 79
References 80
Appendix
LIST OF TABLES
1: Composition and Abundance of pollen grains in honey samples
from study area 54
2: Results of Mean and Ranges of physical content in
honey samples 59
3: Ranges and mean of vitamin content in honey samples 70
4: Antibacterial activities showing zone of
inhabitation at 50mg/ml. 100mg/ml and
150mg/ml in
Nassarawa State 72
5 Antibacterial activities showing zone of
inhabitation at 50mg/ml. 100mg/ml and
150mg/ml in
Plateau State 73
6: Antifungal activities showing zone of inhabitation
at 50mg/ml. 100mg/ml and
150mg/ml in
Nassarawa State 76
7: Antifungal
activities showing zone of inhabitation at 50mg/ml. 100mg/ml and
150mg/ml in
Plateau State 77
LIST OF FIGURES
1. Map showing North Central state of Nigeria 30
2. Means of physical composition of honey
sample from
Nassarawa State and Plateau state. 60
3. Mean of
physical composition (colour intensity) of honey
samples from Nassarawa State and
Plateau state 61
4. Means of
proximate content (moisture contents,
dry matter, carbohydrate) in honey samples 61
5. Means of
proximate content (Ash and crude protein)in honey samples
62
6. Means of Macro nutrient composition of honey
samples 64
7. Means of Macro nutrient composition of honey samples 65
8.
Rangers and Means of Micro element content in honey samples 66
9. Means of sugar
content 68
10. Means of phytochemical contents in
honey samples 69
LIST OF PLATES
1.
Traditional bee keeping
practice using log wood in Bokkos L.G.A of Plateau state 32
2.
Traditional beekeeping
practice mud using house hives in Akwanga L.G.A. In Nasarawa state. 33
3.
Modern beekeeping practice
using Kenyan top bar hives with artificial 33
shade in Karu L.G.A. in
Nasarawa state
33
4.
Modern beekeeping using
Kenyan top bar with natural shade 34
(Tectona grandis stand) in Jos north
L.G.A in Plateau state.
34
5.
Showing honey samples collection
procedures
34
6.
Showing honey samples collection procedures 35
7.
Showing honey bees on identified plant 35
8. (i-ix)
Identified pollen spectra of honey samples from study sites 55
(Nassarawa
State)
9. (i-ix)
Identified pollen spectra of honey samples from study sites 56
(Plateau State)
10. (i-viii)
Identified pollen spectra of honey samples from study sites 103
(Nassarawa and Plateau
State)
LIST OF APPENDICES
1.
Analysis of variance for Pollen
grains
91
2.
Plant species within study area
93
3.
Table (3a-g) Result of Ranges and Means of
physico-chemical composition of
honey
sample from Nassarawa State and Plateau state.
95
4.
Table (4a-f) Result of antimicrobial
activities of honey samples from the study area 103
5. Honey
colour grading chart 105
CHAPTER
1
INTRODUCTION
1.1
BACKGROUND OF THE STUDY
Honeybees provide
pollination for various plant species. Honeybee products are also useful in
food, honey and wax for cosmetic and pharmaceutical industries (Tammy 2008).
Honey collection is a long aged traditional activity throughout most parts of
Africa. Traditional Beekeepers use simple hives made from calabash, clay pot or
hollow wood logs, closed at both ends with a hole as an entrance for the bees.
The empty hives are then placed on trees and become occupied by-passing swarms
and in due course are plundered by bee keepers, the above methods of harvesting
or beekeeping practice have led to a poor yield of low quality honey. However,
now modern beekeepers practice the management of Bees with modern bee hives
like the Kenyan- top bar hives and the langstroth hives which are placed within
apiaries to maximize the goods and
services delivered by the honey bees (Amulen et. al., 2019).
The
benefit of keeping bees are numerous, most people think of the rewards of
harvesting their own honey or the positive impact which the increased
pollination will have on their garden while these are great reasons to have a
beehive does not count out the many other bee products you can harvest from a
hive. The positive impact the bees will have on the extended environment around
you or remarkable adventure of bee keeping (Joanna et. al., 2018).
Honey is a fascinating substance; bees make
honey out of flower nectars for use as a food source which is stored in combs.
Honey does not only taste great but also can be beneficial to those with allergies.
The theory is that since bees collect pollen where you
live, eating the small amounts of pollen present in local honey can actually
immunize the body against the pollen so there is minimal irritation during
allergy season. Honey also have anti-bacterial, and anti-microbial properties
which can be used for wound treatment especially for burns and has also been
used for centuries as skin and hair beautifiers, and recipes for face mask,
shampoos and bath oils among others. The highly industrious bees also create a
variety of other fascinating substances including propolis, bee
pollen and royal jelly. Honeybees also
help in pollination which can make flowers and plants to be more profuse and
fruit and vegetable yields to increase dramatically (Robyn
et. al., 2015). Not only do honey
bees have enormous positive impact on gardens but their nectar gathering range
allows them to also fly outside their range and pollinate an extended area.
Pollination helps the ecosystem to remain diverse and sustainable.
Parasites of diseases on the, other hand, pesticides, have wiped out
most of the native varieties of honeybees. Honeybees belong to the order:
hymenoptera (i.e. insects with membranous wings which also include wasps, sand
fly and ants). Honeybees are members of super family: Apodiaei, family: Apidae,
Sub-family: Apinae, Tribe: Apini, Genus: Apis and can be divided into three (3) branches based on how honey
bees nests are made.
The giantv-open nesting honey
bee, the dwarf single combined honey bees Apis
florae.
The different honey bee species thrive in
extreme habitats such as deserts, rain forest, tundra, but most people only
know Apis mellifera, the agricultural
darling (Crane1980). However, Beekeeping
is a branch of forestry and
agriculture with major concern on effective and efficient pollination service
which is useful and beneficial to man and his ecosystem.
Honeybees also produce honey, beeswax,
pollen, propolis, bee venom, and royal jelly which serve very useful purpose in
various industries. Beekeeping does not involve much human labour; the honey
bees do the real work of collection and conversion of nectar into honey which
influences colony growth and yield (Bradbear 2009).
Furthermore,
the management skills and concept of every bee keeper should be focused on the
interactions of the honey bees and its immediate environment (vegetation belt).
This is because honeybees visit various pasture plants during foraging.
However, climatic condition also affects honeybee activities and invariably
honey yield.
1.2 STATEMENT OF PROBLEM
The continuous increase in human population at
the rate of 3.1-3.2% per annum on fixed area of land has resulted in shortage
of not only food but also of food products, medicinal plant and other raw
materials (Varole et. al., 2019).
Shifting cultivation system used in food production in the study area is no
longer viable because of land shortage. Continuous destruction of forest and
different vegetation zones as a result of incessant demand for land is
affecting honey bee population resulting in decrease in population of honeybee
and invariably pollination of plants, honey yield and loss of gene pools. The
current status / extent of this decrease needs to be investigated in the study
area (the guinea savannah zone in Nigeria) via this research project.
1.3 AIM OF THE STUDY
The aim of this study was to identify some of the plant
species visited by honey bees in the guinea savanna region of Nigeria through
pollen analysis.
1.4 OBJECTIVES OF THE STUDY
The research specifically has the following
objectives,
i. To determine the
composition and frequency of occurrence of pollen grains in various honey
samples in the study area.
ii. To determine pollen spectra
of the study area.
iii. To determine the
physico-chemical and phyto-chemical compositions of the honey samples collected
from study area.
iv. To determine antimicrobial
activities in honey sample from study area
1.5 JUSTIFICATION OF THE STUDY
Beekeeping is practiced all over the world.
However, honey quality as well as quantity varies widely depending on several
factors like floral composition of various vegetation belts and other prevailing
environmental conditions. This study will be of immense benefit to beekeepers
and other forest allied industries. There is need to increase food production
and other raw materials. The increase in availability of information on plant
species visited by honeybees will guide beekeepers on the type of melliferous plants to be protected and
propagated and this will also increase honey yield.
Since there is shortage of land for use in practicing
shifting cultivation, pollination by honeybees becomes one of the best alternatives.
Also there is need to increase honey and other honeybee products which are
useful in various industries such as food, pharmaceutical, cosmetic etc. this
can only be achieved by identifying forage sources of honeybees and increase
their availability.
It will also create direction to health communities on
where to source choice quality honey because of its medicinal benefit to
humans.
1.6
SCOPE OF THE STUDY
The scope of this
study is to identify the pollen composition of honey produced by West African
honeybee (Apis mellifera adansonii)
within the guinea savanna vegetation zones of Plateau State and Nasarawa State
in order for plant species to be protected, propagated and conserved by
beekeepers.
The laboratory
analyses to identify pollen compositions of each honey sample from various
study sites were carried out at Archeology and Anthropology Department of
University of Ibadan, Oyo state using Acetolysis method and while proximate
composition of honey samples was carried out at Plant Pathology Laboratory of National Root Crop Research Institute,
Umudike Abia State.
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