JIMMA UNIVERSITY

COLLEGE OF AGRICULTURE AND VETERINARY MEDICINE

SCHOOL OF VETERINARY MEDICINE

EVALUATION OF HONEY QUALITY AND BEEKEEPING

SYSTEM IN SELLECTED DISTRICT OF HORO GUDURU WOLLEGA ZONE,
OROMIA REGION , WESTERN ETHIOPIA

MSC THESIS

BY
RAJI DUGASA

JUNE 2020
JIMMA, ETHIOPIA
 Jimma University

College of Agriculture and Veterinary Medicine

School of Veterinary Medicine

Evaluation of Honey Quality and BeekeepingSystem In Sellected District of Horo Guduru

Wollega Zone, Oromia Region , Western Ethiopia

A Thesis Submitted to the School of Veterinary Medicine, Jimma University College of

Agriculture and Veterinary Medicine in Partial Fulfillment of the Requirements for the
Degree of Master of Science in Veterinary Public Health

By
Raji Dugasa

Major Advisor:Prof. Tadele Tolosa (PhD)

Co-Advisor:Dr. Tadele Kabeta (DVM, MVPH, Assist. Professor)

June 2020
Jimma, Ethiopia
I
I
DEDICATION

I dedicate this piece of work to my mother, for her committed lives with strong prayers for the piece
and health of our family. I dedicate it also to my beloved wife Beshatu Alemayehu for being
providing support and inspiration to my academic journey.

I
II
BIOGRAPHICAL SKETCH

The author was born in 1990 G.C. from his father Dugasa Yadeta and his mother Dinkinesh Dugasa
in Amuru Wereda, Horo Guduru Wollega zone, Oromia regional State, Ethiopia. He completed his
elementary school in Ejere Goromti elementary school and Amuru primary and secondary school
from 1996 to 2003 and high school education in Fiche high school from 2004to 2005 and
completed his preparatory school in Amuru Preparatory school from 2006 to 2007.

After passing the Ethiopian School Leaving Certificate Examination in 2008, he joined Haramaya
University Faculty of Veterinary Medicine in October, 2008 and graduated with Doctor of
Veterinary Medicine on 12th July, 2012. After he received his DVM degree, he was employed by
Oromia Special Zone of Amara Regional State Agricultural office on Febreury, 2013 as a junior
animal health prevention and quality control expert at Kamissie administrative town, Oromia
special Zone and he served there until June, 2015.

Since June 2015 he was transferred to Horo Guduru Wollega Amuru district Livestock
Development and Fishery Office served at different position as an expert, team leader, and animal
health extension and technology dissemination team coordinator. On October, 2018, he joined
Jimma University College of Agriculture and Veterinary Medicine to pursue his postgraduate
studies in Veterinary Public Health stream from October, 2018 to date.

III
ACKNOWLEDGEMENTS

I wish to thank the Almighty God for giving me the chance to enjoy the fruits of my endeavor. This
work was made possible through the combined efforts and positive attitudes of many kind-hearted
persons all of whom I thank very much. I shall only mention few names among these: I would like
to extend my deepest appreciation and heartfelt thanks to my major and co-advisors, Prof. Tadele
Tolosa and Dr. Tadele Kabeta respectively, for their valuable comments, supports and follow up
from the preparation of the proposal up to the end of the research work. Their careful follow up and
guidance throughout the study period has contributed a lot to cover and complete timely the wider
area coverage of the study. I have deep sense of respect for Prof.Taye Tolamariam for his
constructive and precious suggestions and comments and all staff members of Jimma University
College of Agriculture and Veterinary Medicine, particular School of Veterinary Medicine all my
teachers who taught me various courses and for all their support for research work.

Special thanks should also go to Amuru district and Horo Guduru Wollega Livestock Development
and Fishery zonal Office for offering me the chance to pursue my MSc study. My deepest gratitude
also goes to my hero aunt Tezera Yadeta and his family for nursing me throughout my education
life from under graduate to date. Also, I would like to say thank you to Holeta Bee Research Center
and Wollega University for facilitating the necessary logistic, reagents and laboratory services
especially professional Deressa Kebede for his wonderful during my laboratory work.

No word to speak, no paper to write and no idea to think in my language shall ever express my
deepest sense of gratitude to my lovely wife Beshatu Alemayehu and my child Labsi Raji for
their enormous patience during this long study without hesitating to be alone and holding all
responsibilities.

Lastly, I would like to acknowledge all my precious brother Habtamu Dugasa and friends Drs.
(Yoseph Tilahun, Garoma Desa, Dirriba Oljira and Wakuma Mitiku) for their positive attitudes and
generous idea towards other person. And also every person who has come in to my life directly or
indirectly and inspired me through their presence.

IV
ABBREVIATIONS AND ACRONYMS

AFB American foulbrood

ANOVA Analysis of Variance
ARDO Agricultural and Rural Development Office
BC Before Christ
CAC Codex Alimentarius Commission
CAS Codex Alimentarius Standard
COMESA Common Market for Eastern and Southern Africa
CSA Central Statistical Authorit
DAs Development Agents
Ec Electrical conductivity
EFB European foulbrood
EU European Union
FAO Food and Agricultural Organization
G Conductance in ms
ha hectar
HBRC Holeta Bee Research Center
HGWZLRDFO Horo Guduru Wollega Zone Livestock Resource Development and
Fishery Office
HHs House Holds
HMF Hydroxymethylfurfural
HO Health Office
IHC International Honey Commission
JJWANRO Jardaga Jarte Woreda Agricultural and Natural Resource Office
K cell constant in cm -1
KAP Knowledge, Attitude and Practice
LRDFO Livestock Resource Development and Fishery Office
M1 Weight of dish + ash
M2 weight of dish
Mc Moisture content
Mo weight of Honey
MoA Ministry of Agriculture
mS .cm-1 milli Siemens per centimeter
MS excel Micro Soft excel
NGO Non-Governmental Organization
QSAE Quality Standard Authority of Ethiopia
RFI Radio Frequency Identification
RI Refractive Index
SH Electrical Conductivity of honey solution in mS cm-1
SPSS Statistical package for social science
W Water content in g/100

V
TABLE OF CONTENTS PAGE

DEDICATION................................................................................................................................. I
BIOGRAPHICAL SKETCH ...................................................................................................... III
ACKNOWLEDGEMENTS .........................................................................................................IV
ABBREVIATIONS AND ACRONYMS ...................................................................................... V
TABLE OF CONTENTS .............................................................................................................VI
LIST OF TABLES .................................................................................................................... VIII
LIST OF FIGURES OR ILLUSTRATIONS .............................................................................. X
LIST OF ANNEXES .....................................................................................................................XI
ABSTRACT ................................................................................................................................. XII
1. INTRODUCTION....................................................................................................................... 1
1.2. Objectives .............................................................................................................................. 4
1.2.1. General objective ............................................................................................................. 4
1.2.2. Specific objectives ........................................................................................................... 4
2. LITERATURE REVIEW .......................................................................................................... 5
2.1. People, Bees and Rural Development ................................................................................. 5
2.2. Physico-chemical Property and Quality of Honey ............................................................ 6
2.2.1. Hygroscopicity ................................................................................................................ 8
2.2.2. Viscosity .......................................................................................................................... 8
2.2.3. Density............................................................................................................................. 8
2.2.4. Color ................................................................................................................................ 8
2.2.5. Electrical conductivity (Dry matter of honey) ................................................................ 9
2.2.6. Moisture........................................................................................................................... 9
2.2.7. pH and free acidity = which indicates degree of fermentation...................................... 10
2.2.8. Reducing sugar .............................................................................................................. 10
2.2.9. Total ash ........................................................................................................................ 10
2.2.10.Hydroxymethylfurfural ................................................................................................ 11
2.3. Public Health Benefits of Honey ....................................................................................... 12
2.4. Overview of Beekeeping in Ethiopia................................................................................. 14
2.5. Factors Affecting Beekeeping and Honey Quality in Ethiopia ...................................... 16
2.5.1. Type of flowers ............................................................................................................ 17
VI
 2.5.2. Blending ....................................................................................................................... 17
2.5.3. Storage and heating ...................................................................................................... 18
2.5.4. Agro-chemicals ............................................................................................................ 18
2.5.5. Honey Bee Disease ...................................................................................................... 19
2.5.6. Honey bee predators and pests ..................................................................................... 20
3. MATERIALS AND METHODS ............................................................................................. 21
3.1. Description of the Study Area ........................................................................................... 21
3.2. Study Design ....................................................................................................................... 23
3.3. Sampling Method, Sample Size Determination and Data Collection ............................ 23
3.3.1. Honey sample for honey quality analysis ....................................................................... 24
3.3.1.1. Honey sample size determination and sample types............................................... 24
3.3.1.2. Sample preparation and handling procedure........................................................... 25
3.3.2. Questionnaires................................................................................................................. 27
3.4. Data Analysis ...................................................................................................................... 29
4. RESULTS .................................................................................................................................. 30
4.1. Results of Honey Quality Parameters .............................................................................. 30
4.1.1. Moisture Content ............................................................................................................ 32
4.1.2. Hydroxymethyl furfural (HMF) ...................................................................................... 33
4.1.3. pH.................................................................................................................................... 33
4.1.4. Free acidity...................................................................................................................... 34
4.1.5. Ash content ..................................................................................................................... 34
4.1.6. Electrical conductivity .................................................................................................... 35
4.1.7. Reducing sugar................................................................................................................ 35
4.1.8. Apparent Sucrose ............................................................................................................ 36
4.1.9. Color ............................................................................................................................... 36
4.2. Results of Univariate General Linear Model Analysis of Honey Quality Parameters 37
4.3. Results of Multivarite General Linear Model Analysis of Honey Quality Parameters37
4.4. Correlation between the Quality Parameters of Honey Sampled .................................. 39
4.5. Questionnaire Survey ......................................................................................................... 40
4.5.1. Socio-demographic characteristics and beekeeping system of the respondents ........... 40
4.5.2. Knowledge, attitude and practices of beekeepers on public health benefits of honey .. 42
VII
 4.5.3. Honey bee flora and their flowering season .................................................................. 47
4.5.4. Beekeeping challenges in the study area ....................................................................... 48
5. DISCUSSION ............................................................................................................................ 50
6. CONCLUSION AND RECOMMENDATIONS .................................................................... 60
8. ANNEXES ................................................................................................................................. 71

VIII
LIST OF TABLES PAGES

Table 1. Traditional and ayurveda recipes with honey ................................................................ 14

Table 2. Summary of purposively selected kebeles and collected honey samples ...................... 25

Table 3. Total selected kebeles and beekeepers for questionnaire survey ................................... 29

Table 4. All mean results of honey samples collected from the study area ................................. 31

Table 5.Mean results of honey samples collected from different Agro-ecology and hive types . 32

Table 6. Multivarite analysis between sources............................................................................ 38

Table 7. Multivarite analysis among agro-ecologies .................................................................. 38

Table 8. Pearson correlation coefficients among the analyzed parameters .................................. 39

Table 9. Socio-demographic indicators and beekeeping system of respondents (N= 125) ......... 41

Table 10.Hive types owned by beekeepers of respondents among agro-ecologies ..................... 42

Table 11.Knowledge, attitude and practices of beekeepers on public health benefits of honey .. 44

Table 12.Summary of medicinal value of honey in the study area ............................................. 46

Table 13.Major honey bee floras and blooming season in the study area ................................... 48

Table 14.Major constraints of beekeeping identified in the study area (N= 125) ........................ 49

IX
LIST OF FIGURES OR ILLUSTRATIONS PAGES

Figure 1.HMF effects on honey bee and human health…………………………………………..12

Figure 2. Location map of study area……………………………………………………………..21

Figure 3. Prepared and labeled honey samples for laboratory analysis…………………………...26

Figure 4. Some flowering plants in the study area………………………………………………..47

X
LIST OF ANNEXES PAGES

Annex 1. Sample collecting sheet ..................................................................................................... 71

Annex 2. Questionnaire recording sheet ........................................................................................... 71

Annex 3. Procedure for honey moisture determination..................................................................... 72

Annex 4. Conversion factor for the estimation of moisture content from refractive index data ...... 73

Annex 5. Procedures for hydroxyl-methyl furfural ........................................................................... 74

Annex 6. Dilution of sample and reference solutions carried for estimation of HMF ...................... 74

Annex 7. Procedure for Acidity and PH ............................................................................................ 75

Annex 8. Procedures for determining ash content............................................................................. 76

Annex 9. Procedure of determining reducing sugar and sucrose by HPLC Method ........................ 77

Annex 10. Procedures for color determination................................................................................. 79

Annex 11. USDA color standard ....................................................................................................... 79

Annex 13. Results of Univariable GLM analysis of honey samples parameters ........................... 82

Annex 14. Quality of honey sample of the study area as compared to QSAE, EU and CAC: ........ 83

Annex 15. Questionnaires used in the study ..................................................................................... 84

XI
ABSTRACT

Honey is one of the oldest sweetening foods, has medicinal, nutritional and economic importance.
The present work has the aim of evaluating honey quality and assessing knowledge, attitude and
practices (KAP) of beekeepers by using 24 honey samples and 125 selected beekeepers in Amuru
district of Oromia region, west Ethiopia. Of the total of 24 honey samples, of which 18 samples
were purposively taken from selected six kebeles from farm gates of beekeepers and 6 samples
from honey retailers. The main analyzed parameters were Moisture Content (MC), Hydroxymethylf
urfuran (HMF), pH, Free Acidity (FA), Ash, Electrical Conductivity (EC), Reducing Sugar,
Sucrose and Color. The analysis was done in Holeta Bee Research Center. A total of 125
respondents were interviewed and field observation was made used as tool of data collection. The
laboratory result and the survey were analyzed by SPSS version.23 by using GLM, and using
descriptive analysis method respectively. The overall mean ± SD of MC, HMF, pH, FA, ash, EC,
RS, Sucrose and Color of honey analyzed were; 20.43 ± 1.32 %, 16.39 ± 2.68 mg/kg, 3.92 ± 0.07,
36.67 ± 2.13 meq/kg, 0.25 ± 0.14, 0.68 ± 0.3 %, 73.08 ± 0.92 %, 1.80 ± 0.35 % and 103.75 ± 2.89
mm respectively. The value of moisture, HMF and FA of honey significantly (p < 0.05) influenced
by agro-ecologies and sources. pH and Color values of honey from the retailer was significantly
higher (p < 0.05) than the honey collected from beekeepers. There was no significant difference (p
> 0.05) revealed among hive types of all parameters analyzed. The moisture value of honey mainly
declared significantly correlated with free acidity with r = 0 .63**, (p < 0.01) and strong positive
correlation between HMF and EC with the highest r= 0.77 **, (P < .01) is observed. Beekeeping
mainly practiced by males 87.20%. The range of age of respondents were from 25 to 65 years old
mostly. 89.60% knew the health benefits of honey and practiced to treat many of the human and
animal diseases. Presence of honey bee flora, and bee colony considered to be an essential indicato
r for potentialities of the beekeeping of the area. However, pests and predators, indiscriminate agroc
hemicals and diseases,were the major identified beekeeping constraints. It can be concluded that
most results of honey quality analyzed on honey produced in the area is of good quality and can
meet the national and international standards. Beekeepers KAP study indicates honey has a public
health benefits which could be a great contribution for the development of modern
medicine. Therefore, strong efforts have to be made to promote this indigenous knowledge and
practices of beekeepers on honey health benefits and improve beekeeping system through extension
Key words: Beekeepers, Honey quality, Indigenous, KAP, Physicochemical, Retailer
XII
1. INTRODUCTION

Honey is one of nature‟s wonders, of the oldest sweetening foods, has medicinal, nutritional and
economic importance(Abeshu and Geleta, 2016).It is certainly the only sweetening agent that can
be used by humans without processing (Yadata, 2014). History has revealed that humans had
used bee products such as honey for thousands of years in all societies worldwide (Al-Waili et
al., 2012). The method of trapping a colony of bees and then taking them to home yard, allowing
them to rear and multiply, prepare honey and beeswax is known as beekeeping (Seeley,
2019).Beekeeping seems as old as time itself and no one knows exactly when and where it was
started. However, it is believed that, primitive man, may be even Adam and Eve, harvested
honey from bee nests in hollow trees and rock crevices (Fissures) and is portrayed in many rock
paintings in Africa and Europe (Wallace, 2007).

Honey has been used as a food and medical product since the earliest times. It is a natural
substance produced by honeybees, Apis mellifera, from the nectar of blossoms or from exudates
of trees and plants giving nectar honeys or honeydews, respectively. As the only available
natural sweetener, honey was an important food for Homo sapiens from his very beginnings.
Indeed, the relationship between bees and man started as early as the Stone Age (Alvarez et al.,
2010).Honey is the sweet and viscid fluid, contains significant amounts of mineral, vitamins, and
enzymes (Darko et al., 2017). With respect to carbohydrates, honey is mainly fructose (about
38.5%) and glucose (about 31.0%) (Blasco et al., 2011) and other sugars are present as traces,
depending on floral origin. Honey composition varies depending on its floral, geographical and
entomological sources. In addition, external features such as seasonal and environmental factors
honey processing, and storage time and conditions have crucial effects on honey‟s composition
(Gidamis et al., 2004).

Amongst the factors that most influence quality is high temperature, length of storage and
moisture content greater than 21%. They lead to fermentation, high levels of
Hydroxymethylfurfural (HMF), loss of enzymatic activity, changes in flavor, darkening and
microbial growth (Kinati et al., 2011).The quality of honey relied to a great extent on the art of
the producer in storing and blending the product. In marketing of honey, consumers should have
confidence that they are getting good quality for what they are paying so that the country able to

1
earn foreign currency to revamp the national economy (Getachew et al., 2014).Agricultural
contamination with pesticides and antibiotics is a challenging problem that needs to be fully
addressed. Honey, are widely consumed as food and medicine and their contamination may carry
serious health hazards. Pesticide residues cause genetic mutations and cellular degradation and
presence of antibiotics might increase resistant human or animal's pathogens (Al-Waili et al.,
2012). Due to continuous expansion of the world honey market, the importance of apiculture as
an industry has also grown. Composition and quality criteria of honey are defined by the Codex
Alimentarius standard (Souza et al., 2006) and the EU Honey Directive (Fallico et al., 2006)
which state that honey should not have any ingredients added; no particular constituent can be
removed from it; it does not have any objectionable matter, flavor, aroma or taint absorbed from
foreign matter during processing and storage.; and it should not be heated or processed
(Bogdanov et al., 2002).

Honeybees are of critical importance in Africa for both ecological and economic reasons. Their
contribution to floral biodiversity and conservation, by virtue of their pollination of indigenous
flowering plants is unknown, but certain to be considerable. Economically, honeybees are critical
for the pollination of a host of commercial crop plants as well as being the source of energy and
livelihood for many thousands of mostly small-scale beekeepers (Allsopp, 2004). In African
developing countries agricultural production is expected to become increasingly reliant on
pollinator services. However, in response to the increasing challenge of providing food security
in sub-Saharan Africa, farmers have been simultaneously encouraged to adopt intensive
agricultural practices often characterized by widespread use of pesticides as foliar sprays and
seed coatings meaning service provision by bees is contingent upon their ability (Power, 2010).
Food security is not only a matterof producing grains but also the financial power to pay for the
purchase of grain (Caplan, 2002).

Ethiopia occupies the major part of the Horn of Africa. The country covers approximately 1.11
million square kilometers and it is a country of great geographical and climatic diversity, with
varied ecological conditions (Froehlich and Siebrits, 2019).Ethiopia, a potential beekeeping
giant. In an Abyssinian grain-market, many honey bees were observed collecting from open
sacks of shirro (Cicer arientinum) as a pollen substitute. Usage of honey for making “tej”, and
for selling (Hussein, 2001).
2
In Ethiopia beekeeping sub-sector has been an integral part of agriculture. It has been
contributing to the household income and poverty alleviation and national economy through
export. Ethiopia has huge apicultural resources that made it the leading honey and beeswax
producer in Africa (Fikru, 2015). Beekeeping is a long standing practice in Ethiopia and it
accounts 1.3 % of agricultural GDP (Demisew, 2016). According to FAO (2009), report 45,300
metric tons of honey is produced per annum in Ethiopia makes the country to rank first honey
producer in Africa and ninth in the world. However, the majority of honey is crude and poorly
managed. In Ethiopian, only about 10% of the honey produced in the country is consumed by the
beekeeping households. The remaining 90% is sold for income generation and of this amount, it
is estimated that 80% is used for tej brewing (Legesse, 2014).

1.1. Statement of the Problems

Beekeeping activities in Ethiopia is mainly one of income generating economic activities to

subsistence farmer. However, constrained by inability in the transformation, lack of sufficient
awareness on beekeeping practice, promotion, scaling up to rapid growth, lack of commercial
beekeeping development and beekeeping technology, limited credit supply, quality issue, lack of
market access, research and information transmissions (Winberg, 2011).The reports from
different parts of Ethiopia are indicating that extreme use of herbicides, pesticides and insecticide
chemicals are increasing. Studies on the physicochemical and quality analysis have been carried
out in some parts of Ethiopia by different researchers(Berhe et al., 2016). The quantity and
quality of Ethiopian honey in generally poor, as 95% of beekeepers follow traditional method of
beekeeping practice with no improved techniques or technology (Beyene et al., 2014). Honey is
of good quality as long as it is in the hive, but faulty handling from the time of its harvest until it
reaches to market is responsible for its inferior quality. Several factors have contributed to its low
quality among which high moisture content is the major quality problem in Ethiopia. Harvesting
unripen honey, unsuitable honey storage container and storage places also attribute to high
moisture content (Shunkute et al., 2012).

Oromiya, Amahara, Southern National Nationalities and People,Tigray and Benshangul are the
major honey producing regions in Ethiopia, with production quantities of 21,403.404tons, 10,834
tons, 9,471.625tons and 3,293.394tons and 2,231.380 tons respectively (CSA, 2017/2018). In

3
Oromia region beekeeping is also a very long-standing practice in the farming communities and
it plays a significant role as a source of additional cash income and nutrition for many
subsistence farmer beekeepers. It is an integral part of the smallholder farming system. The
natural vegetation coverage is relatively high, as a result in this region the honeybee population
is dense and production is relatively high. Besides, the beekeeping potentiality of the region, it is
partly attributed to the various cultivated oil crops, pulse and field flowers, which are very
important, source of forage (Ambaw and Teklehaimanot, 2018).

Amuru district is found in Horo Guduru Wollega Zone, Oromia region, with suitable agro-
ecology that makes the district potential for honey production. The nature of diversified
flowering plant species and agro-climatic conditions has enabled the area to sustain a number of
honeybee colonies.Despite the potentiality of the areas and large volume of honey production,
there is no scientific research which has been done to quantify and document the actual
properties and quality of honey, beekeepers Knowledge, Attitudes and Practices (KAP).
Therefore, this study has been conducted with the following objectives:

1.2. Objectives

1.2.1. General objective

The aim of the study was to evaluate honey quality, and assess KAP of beekeepers in the
study area.
1.2.2. Specific objectives
To evaluate honey quality
 To assess beekeepers KAP on health benefits of honey
To assess beekeeping system and managements

4
2. LITERATURE REVIEW

2.1. People, Bees and Rural Development

Since ancient times, people have been drawn to the study of bees. Bees are spritely creatures that
move about on pleasant bright days and visit pretty flowers (Genise, 2017).Honeybees have the
capacity to live together harmoniously in a nest, foraging nectar and pollen grains from
flowering plants to satisfy the foodneeds of the colony members in the nest. Honey bees are the
most usefuland friendly insects to all living things on earth. Such insects that live and work
together for their nest activities in an area selected are called social insects (Seeley, 2010). Bees
are sensitive indicators of an intact environment, and as essential and persistent shapers of the
environment, have a significance that cannot be estimated highly enough. Honey bees are very
important for the maintenance of biodiversity (Judova et al., 2016).

Moreover, bees are important pollinators of both natural vegetation and crops, and certain kinds
of bees make useful products, especially honey and wax. We are social animals; some bees are
also social. Their interactions and communications, which make their colonial life function, have
long been matters of interest; we wonder how a tiny brain can react appropriately to societal
problems similar to those faced by other social animals, such as humans. For a biologist or
natural historian, bees are also fascinating because of their many adaptations to diverse flowers;
their ability to find food and nesting materials and carry them over great distances back to a nest;
their ability to remember where resources were found and return to them; their architectural
devices, which permit food storage, for example, in warm, moist soil full of bacteria and fungi;
and their ability to rob the nests of others, some species having become obligate robbers and
others cuckoo like parasites. These are only a few of the interesting things that bees do
(Michener, 2000).

Human experience of beekeeping started 5 million years ago from the earliest times in which
presumably the honeybees coming to existence. The object of exploitation for their by the
ancestors of mankind who appeared 1.8 million years ago, in a manner similar to the one by
which chimpanzees today lick honey of tree twigs by inserting them into wild bee beehives
(Oldroyd and Wongsiri ,2009).

5
Beekeeping is of pivotal importance, combining the wide economical aspect of honey production
and the important ecological services provided by honeybees (Zoccali et al., 2017).

Rural development aims to help people manage their livelihoods better through sustainable use
of the available resources. It provides them with greater social and economic power by offering
them opportunities to work in line with their capacity, without hampering the eco-services
provided by their environment (Ahmad et al., 2007). Beekeeping and honey hunting have been
practiced by different societies since ancient times and have always been linked to development.
„Honey hunting‟ – collecting honey from wild bee colonies – is an ancient practice as shown, for
example, in cave paintings dating back to 11,000 BC found in Madhya Pradesh, India (Roué et
al., 2015).

Beekeeping is the establishment and tending of colonies of social bees of any species. Most of
the world's beekeeping is done with Api mellifera. Beekeeping is done mainly to produce honey,
but there are also other specialized types of operations. These include the rearing of queens or
package bees for other beekeepers that are producing honey. Another type of beekeeping
provides colonies of bees to pollinate crops, since in many areas of large-scale agriculture; the
native pollinators have been destroyed. Each type of beekeeping requires the management of
colonies to stimulate the bees to do what the beekeeper wants-for instance, to rear younger house
bees to produce royal jelly or more foragers to pollinate crops. So it is essential that hives and
frames have standard dimensions and that an accessory is used to ensure that frames are always
exactly the correct distance apart (Crane, 2009).

2.2. Physico-chemical Property and Quality of Honey

The study of the physical and chemical properties of honey has increased in recent years because
these parameters are important for the certification process that determines honey quality
(Farooq and Maqbool, 2008). Honey features vary depending on the botanical source and
geographical origin, as well as climatic, processing and storage conditions. Honey is mainly
composed of carbohydrates and water, parameters that influence its shelf life and some of its
properties, including color, flavor, density, viscosity, hygroscopicity, and crystallization. Honey
also contains small amounts of other components, such as nitrogen compounds, organic acids,

6
minerals, vitamins, Maillard reaction products, volatile compounds, and several bioactive
substances that affect sensory and physical characteristics, as well as biological potential (Da
silva et al., 2016).

The quality of honey is normally assessed by physico-chemical test of its ingredients. These
ingredients have substantial insinuations (hints) on honey industry as they influence the storage
quality, granulation, texture, flavors and nutritional and medicinal value of the product.
Internationally, certain constituents are proposed as quality criteria for honey and these include,
but not limited to, moisture level, electrical conductivity, reducing sugars, amount of fructose
and glucose, concentration of sucrose, free acidity, total acidity, hydroxymethylfurfurale (HMF)
and proline content. The magnitude of these physico-chemical properties of honey could be
influenced, among others, by the type of storage container used. In this regard, it is assumed that
the physio-chemical properties of honey can change with time and kinds of storage containers
(Gebremeskel, 2015).Chemical composition of honey varies depending on plant source, season
and production methods. Storage conditions may also influence final composition, with the
proportion of disaccharides increasing overtime. Amongst the factors that most influence quality
is high temperature, length of storage and moisture content greater than 21%. They lead to
fermentation, high levels of Hydroxymethylfurfural (HMF), loss of enzymatic activity, changes
in flavor, darkening and microbial growth (Kinati et al., 2011).

Quality of honey is mainly determined by its sensorial, chemical, physical and microbiological
characteristics. Pollen profile, color, moisture content, ash, acidity, electrical conductivity, pH,
reducing sugars, apparent sucrose and HMF were the parameters analyzed in each honey sample
(Gomes et al., 2010).A quality product will go a long way in developing the confidence that
encourages return, customers and efficient production of a product to any marketing scheme
(Getachow et al., 2014).External factors like climate, harvesting conditions and storage can also
influence it (Belie, 2009). Inappropriate materials used for honey handling, careless storage
conditions of honey leads to reduce its quality (Mitikie, 2017). The quality and properties of
honey are also related to honey maturity, the production methods, climatic conditions, processing
and storage conditions as well as nectar sources of the honey (Gobessa et al., 2012).

7
2.2.1. Hygroscopicity

The strong hygroscopic character of honey is important both in processing, storage and for final
use. Because of this character it easily absorbs moisture from the air. Thus, in areas with a very
high humidity it can be difficult to produce good quality honey of sufficiently low water content,
which can be measured using a gadget called refracto meter. Different researches show that
normal honey with a water content of less than 18.3 % or less will absorb moisture from the air if
a relative humidity is above 60% (Crane,1996). The moisture content of honey should not be
more than 20% (Ball, 2007).

2.2.2. Viscosity

Viscosity of nectar- and honey-thick liquids measured at a typical serving temperature. Serving
temperature results are contrasted with viscosity measurements collected at room temperature,
showing variable thickening patterns especially related to the type of thickening agent (Garcia et
al., 2008). Freshly extracted honey is a viscous liquid. Its viscosity depends on a large variety of
substances and therefore varies with its composition and particularly with its water content.
Viscosity is an important technical parameter during honey processing, because it reduce honey
flow during extraction, pupping, settling, filtration, mixing and bottling (Olaitan et al., 2007).

2.2.3. Density

Another physical characteristic of practical importance is density. Honey density, expressed as

specific gravity, is greater than water density, but it also depends on the water content of the
honey. Because of the variation in density it is sometimes possible to observe distinct
stratification of honey in large storage tanks. The high water content (less dense) honey settles
above the denser, drier honey. At the temperature of 20 °C, density of honey typically ranges
between 1.38 and 1.45 kg/l (Suliman, 2017).

2.2.4. Color

Honey is color graded into light, amber, and dark categories which do not really have any
bearing on quality. Some of the most distinctively and strongly flavored honey variety such as

8
basswood is very light, while very mild and pleasant honeys such as tulip poplar can be quite
dark. Honey color is measured on the P fund Scale in millimeters. While it is not an indicator of
honey quality and there are exceptions to the rule, generally speaking, the darker color the honey,
the higher its mineral contents, the pH readings, and the aroma/flavor levels. Minerals such as
potassium, chlorine, sulfur, iron, manganese, magnesium, and sodium have been found to be
much higher in darker honeys (Mahmoud, 2012).

2.2.5. Electrical conductivity (Dry matter of honey)

The electrical conductivity of honey is defined as that of a 20% weight in volume solution in
water at 20 0C, where the 20% refers to honey dry matter. The result is expressed in
milliSiemens per centimetre (mS.cm-1).The method is valid for the determination of the
electrical conductivity of honey in the range 0.1 - 3 mS.cm-1.This measurement depends on the
ash and acid contents of honey: the higher their content, the higher the resulting conductivity
(Bogdanov et al., 2002). It is a very easy and quick method, needing only inexpensive
instrumentation. The conductivity is a good criterion of the botanical origin of honey and thus is
very often used in routine honey control. A lower limit has been proposed for blossom than for
honeydew honeys. Exceptions have to be made for some blossom honeys (Bogdanov and Peter,
2002).

2.2.6. Moisture

The moisture content is the most essential quality component of honey, because the rate of
fermentation, its shelf life span and processing characteristics are greatly determined by the
amount of moisture content. The different moisture content of honey depends on harvesting
season, the degree of maturity that honey reached in the hive, type of hive used, environmental
temperature and moisture content of original plant (Gebremedhin et al., 2013). Moisture content
of honey can naturally be as low as 13 % or as high as 23 % depending on the source of the
honey, climatic conditions and other factors (Meixner, 2010).

9
2.2.7. pH and free acidity= which indicates degree of fermentation

Honey pH depends on both the ionized acids of this food and mineral elements and influences
microorganism‟s development, enzymatic activity and texture, among other properties. Honey
typically has a pH in the range of 3.3–5.6. The natural acidity of honey inhibits growth of many
pathogenic bacteria whose minimum tolerated pH is in the range of 4.0–4.5. These two
properties of honey can influence honey stability and its storage conditions and also they give
some information on honey origin (Derebaşıet al., 2014). The high acidity of honey is an
indication of the fermentation of sugars presents in the honey into organic acid particularly the
gluconic acid and the inorganic ions such as phosphate and chloride. According to Bogdanov et
al. (2009) these honey fermentation results are responsible for two important characteristics of
honey: flavor and stability against microbial spoilage (El Sohaimy et al., 2015). The acidity of
honey developed due to the presence of organic acids. The value of honeys acidity, lower than 50
meq/kg of honey, means that honeys will not be fermented (Gebremeskel, 2015).

2.2.8. Reducing sugar

Apparent reducing sugars‟ are defined as those sugars which reduce a Fehling‟s reagent under
the conditions specified. „Apparent sucrose‟ is defined as 0.95 of the difference in „apparent
reducing sugars‟ before and after the prescribed hydrolysis procedure. This method is a
modification of the Lane and Eynon procedure, involving the reduction of Soxhlet‟s
modification of Fehling‟s solution by titration at boiling point against a solution of reducing
sugars in honey using methylene blue as an internal indicator. The difference in concentrations
of invert sugar is multiplied by 0.95 to give the apparent sucrose content. This method is based
on the original method of Lane and Eynon and is also used in the Codex Alimentarius standard
(Bogdanov et al., 2002).

2.2.9. Total ash

The ash content is a quality criterion for honey origin, the blossom honeys having lower ash
content than the honeydew ones. The amount of minerals present in honey does not significantly
contribute to the dietary recommendations. This method will probably be replaced by the faster
and easier conductivity measurement (Downey et al., 2005).
10
Mineral content in honey is generally low, ranging between 0.02 and 0.3% in blossom honeys,
while in honeydew honeys can reach 1% of the total. It is influenced by soil and climatic
conditions, as well as the chemical composition of nectar that varies according to the different
botanical sources involved in honey formation. Variations can also be related to harvesting,
beekeeping techniques and the material collected by the bees during foraging on flowers.
Minerals are absorbed in their salt forms dissolved in water, moving from the roots to the plant
sap and then being pumped to the nectar or honeydew and pollen (Ouchemoukh et al., 2007).
The most important minerals found in honeys are potassium, sodium, calcium and magnesium.
Less abundant elements are iron, copper, manganese, chlorine and in minor quantities trace
elements such as boron, phosphorus, sulfur, silicon, bare and nickel, among others. Potassium is
the main one, standing for 80% of the total, as a result of its quick secretion by nectar sources
(Machado et al., 2017).

2.2.10.Hydroxymethylfurfural

Hydroxymethylfurfural (HMF) is a furanic compound produced by sugar degradation, from

dehydration of hexoses in acidic medium and to a lesser extent, as an intermediate in the
Maillard reactions. HMF is a parameter of honey freshness, since it is absent or present in trace
amounts in fresh honeys. High values of HMF are naturally present in honeys from warm climate
areas, such as tropical and subtropical. HMF concentration increases during honey processing by
heat treatment, and also by adulteration with commercial sugars and throughout storage (Shapla
et al., 2018). HMF content is also affected by the use of metallic containers, pH, bee species and
botanical source. In addition to processing, storage conditions affect the formation of HMF, and
HMF has become a suitable indicator of honey quality. Its concentration increases when honey is
heated and is in its storage (Mehryar et al., 2013).

If you expose samples of honey at the temperature of 60°C for a longer period of time, a
significant increase in the concentration of HMF is present. Noticeable increase of HMF
concentration is observed by heating the samples at 90°C for 60 minutes where the results show
that the average concentration of HMF was 48.8 mg/kg. Therefore, the content of HMF in honey
is an important parameter in determining the quality of honey, his age, antioxidant activity, as
well as the loss of its nutritional value (Kesic et al., 2017).
11
Figure 1. HMF effects on honey bee and human health

Source: Shaplaet al., 2018.

2.3. Public Health Benefits of Honey

Honey is highly nutritious, it has traces of minerals and vitamins not to mention the antioxidants
which destroy free radicals and delay ageing. In short, it is a safe and wholesome food for old,
children and adults. Honey is a plant by-product and used medicinally around the world (
Pollan, 2008). Honey is also an energizer, helping workers and athletes overcome fatigue and
regain energy. Children, young and old can alike take honey, without worrying any side effects.
Honey is a multivitamin tonic, has antibacterial properties and has antioxidants. Asthmatic
persons can also benefit from taking honey every day. Ayurveda acknowledges honey as a
wonder medicine capable of providing longevity. Osteoporosis is another condition, which can
be prevented by taking honey regularly. Modern researches have shore up the wonderful effects
of honey, proving honey to be effective against advanced cases of stomach and bone cancer.

12
Taking one tablespoon honey with one teaspoon cinnamon powder three times a day and cancer
symptoms receded in one month (Kumar et al., 2010).

Honey has been used as medicine in many cultures for a long time. However, it has limited use
in medicine due to lack of scientific report. In recent days, honey is becoming acceptable as a
reputable and effective therapeutic agent. Its beneficial role has been endorsed to its
antimicrobial, anti-inflammatory and anti-oxidant activities as well as boosting of the immune
system. Honey has proven antimicrobial activity (Dureja et al., 2003). Honey inhibits a broad
spectrum of bacterial species. The alcohol extracts of honey exhibit an effect to array of bacterial
species including aerobes and anaerobes, Gram positives, and Gram negatives. Honey has
powerful antimicrobial effects against pathogenic and nonpathogenic micro-organisms (yeasts
and fungi), even against those that developed resistance to many antibiotics. The antimicrobial
effects could be bacteriostatic or bactericidal depending on the concentration that is used
(Abeshu, 2016).

Furthermore, honey is used as a remedy for diarrhea and gastroenteritis at a concentration of 5%

(v/v). Honey reduces the activities of cyclooxygenase-1 and cyclooxygenase-2, thus showing
anti-inflammatory effects and demonstrates immune modulatory activities. Ingestion of diluted
natural honey showed reduction effect on concentrations of prostaglandins such as prostaglandin
E2, prostaglandin F2α, and thromboxane B2 in plasma of normal individuals. Anti-inflammatory
activity of honey was as effective as prednisolone, reference drug. Further, honey has an anti-
inflammatory action free from adverse side effects such as suppression of immune response and
tissue growth, formation of ulcers in stomach (Nweze et al., 2019).

13
Table 1. Traditional and ayurveda recipes with honey

Diseases Recipes
Indigestion Ginger (Zingiber officinale) juice with honey, Lemon (Citrus limon)
juice with honey with Navaratna kalka Roasted cumin seeds powder
with bee honey, Roasted cloves powder with bee honey.
Peptic ulcers Paste prepared with Vishnukranti, honey, ghee and sugar
Diarrheawith vomiting Decoction prepared by Bark of Beli (Aegle marmelos) root and
internal part of the mango seed with honey.
Cough Adathoda (Adatoda vesica) svarasa with bee honey, Decoction of
Adathoda, Elabatu (Solanum indicum) roots and Rasakinda
(Tinospora cordifolia) with honey, Powder of vibitaka (Terminalia
bellirica) 10 g mixed with bee honey cure Asthma and cough
immediately.
Asthma Most of the rasa preparations “Buddharaja kalka” - prescribed with
juice of Ambuldodam (Citrus aurantium), ginger, honey and the drug
“Svasakuthara rasa” is given with honey.
Hiccup Curd with bee honey, Ash of peacock feathers with honey, Rasa
preparation “Arogyavardhanavati” with honey.
Anorexia Pomegranate juice and rock salt with honey

Source: Liyanage and Horadugoda, 2017.

2.4. Overview of Beekeeping in Ethiopia

Beekeeping is a long standing practice in Ethiopia and it accounts 1.3 % of agricultural GDP.
Currently one out of ten rural households keep honeybees and the activity make a substantial
contribution to rural income generation. Ethiopia is the leading honey producer in Africa and
one of the top ten worldwide (Demisew, 2016). Although difficult to establish a time reference
when beekeeping was started in Ethiopia, it may date 5000 years back and the Hieroglyphs of
ancient Egypt refers to Abyssinia (the former name of Ethiopia) as the source of honey and
beeswax.

14
Thus Abyssinia has been known for its beeswax export for centuries during when other items
were not exportable (Cannon, 2009). Ethiopia is one of the poorest and most food-insecure
countries in the world. It is primarily a net exporter of agricultural products, with 85 percent of
its population employed in agriculture. Ethiopian agriculture contributes more than 45 percent to
the nation‟s gross domestic product (GDP) and significantly affects the country‟s export trade
(Dixon et al., 2001).

Ethiopia is the first African countries with huge honey and beeswax producer and having rich
plant biodiversity for bee forage. The ideal climatic conditions and diversity of floral resources
allow the country to sustain around10 million honeybee colonies, of which 7 million are kept in
local beehives by farmers and the remaining, exist in the forests as wild colonies (Ambaw and
Teklehaimanot, 2018).Ethiopia is famous for its notable variation of agro-climatic conditions and
biodiversity which favored the existence of diversified honeybee flora and vast number of
honeybee colonies. The country has the largest bee population in Africa with over 10 million bee
colonies (Bekele, 2018). Beekeeping is a traditionally well-established household activity in
almost all parts of Ethiopia. Ethiopian honey production is characterized by the widespread use
of traditional technology resulting in relatively low honey production and poor quality harvested
when compared to the potential honey yields and quality gains associated with modern beehives
(Arzaga et al., 2017). Considering its importance to the overall growth of the agricultural sector,
the government of Ethiopia gives special emphasis to enhance honey production through the
promotion and expansion of beekeeping activities (Bewket et al., 2015).

2.4.1. Role of beekeeping in Ethiopian

Unlike developing countries, in the most developed nations, the primary objective of keeping
honeybees is for the pollination of various plants. The secondary use of keeping bees is simply
for the production of bee‟s products namely honey and beeswax. In Ethiopia and other
developing countries, the basic purpose of beekeeping is to produce honey and beeswax to get
better income and to assure food security (Masehela, 2017). The beekeeping sub-sector has been
an integral part of agriculture in Ethiopia. It has been contributing to the household income and
poverty alleviation and national economy through export. The country has huge apicultural
resources that made it the leading honey and beeswax producer in Africa. Moreover, Ethiopia is
15
a country where apicultural research is being conducted in a coordinated manner under the
national agricultural research system. Hence, a lot of information has been gathered on different
aspects of the beekeeping. It has been revealed that the country‟s beekeeping subsector is mainly
practiced using traditional basket hives with low productivity. However, attempts by various
investigators and development actors showed that both the production and quality can be
improved in terms of transforming the beekeeping system, processing and marketing (Gemechu,
2014).

Beekeeping in Ethiopia plays an important role in income generators for beekeepers (farmers). In
the country, an average of 420 million ETH. Birr or 35 million $USD obtained annually from the
sale of honey. Honey production of the country meets beverage requirement of the urban and
rural population. It is also demanded for its nutritional and medicinal values. The others hive
products beeswax; royal jelly; propolis and bees venom have high demand globally (Yirga and
Ftwi, 2010).

2.5. Factors Affecting Beekeeping and Honey Quality in Ethiopia

Thirty-five percent of global production from crops including at least 800 cultivated plants
depends on animal pollination. The transformation of agriculture in the past half-century has
triggered a decline in bees and other insect pollinators (Nicholls and Altieri, 2013). Problems and
dangers confront the long-time survival of beekeeping as a profitable agricultural enterprise, and
changing agricultural and land-use practices threaten the survival of adequate numbers of bees
required to pollinate some 90 crops or more. As human population increases, houses, factories,
and highways replace open fields of honey and pollen plants. Clean cultivation of farmland and
large-scale monoculture reduce the sequence of wild plants needed to provide bee food
throughout the season (Kourí, 2004). Honey is produced primarily from floral nectars, and
fructose and glucose are the major components. The chemical composition of honey varies
depending on plant, season, production methods, and storage conditions (Da Silva et al., 2016).
Ethiopia has immense natural resources for beekeeping activity. However, like any other
livestock sector, this sub Sector has been seriously devastated by complicated constraints.

16
The prevailing production constraints in the beekeeping sub sector of the country would vary
depending on the agro ecology of the areas where the activities is carried out. The major
constraints that affect beekeeping sub-sector in Ethiopia are: lack of beekeeping knowledge,
shortage of skills man power, shortage of bee equipment‟s, pests and predators, pesticide threat,
poor infrastructure development and lack of research extension (Beyene et al., 2014).

Honey bees are verysensitive organisms, and their vulnerability toward chemical pollution is a
priorityenvironmental issue. The “hide model” is able to consider different contamination
pathways form inside the hive via pesticide treatments against bee pests or from outsideby means
of the eventual contamination present in nectar, pollen, resin, water, air, orvegetation. Radio
frequency identification (RFI) systems are noncontact identificationdevices now commonly used
in numerous domains, including the study of insectbehaviors, like when faced with pollutants
(Judova et al., 2016).The existing production constraints in the beekeeping development of
Ethiopia are complex and to a large extent vary between agro-ecological zones and production
systems (Ejigu et al., 2009). Most research reports revealed that the pests and predators, shortage
of bee forage, lack of skill and knowledge, low level of technology and honey bee disease, agro-
chemical, are the top major constraints in most part of the country (Abebe and Puskur, 2011;
Godifey, 2015).

2.5.1. Type of flowers

According to the Honey Research Center at the University of Waikato, New Zealand, there is not
enough evidence to draw conclusions on the properties of honey, especially the antimicrobial
properties, based on the type of flowers used for its production. However, extensive research has
been carried out on the honeydew variety obtained from the conifer forests in the central
European mountains and the manuka variety obtained from New Zealand. The above-mentioned
honeydew kind has been found to have a high microbial activity while the manuka kind has been
found to have high non-peroxide activity (Saranraj et al., 2016).

2.5.2. Blending

It is also believed that polyfloral honey (which is obtained from more than one flower) provides
more benefits than monofloral. Hence, many companies sell blended honey as it offers the
17
benefits from a variety and is, therefore, considered to be healthier than non-blended honey
(MUT, 2018).

2.5.3. Storage and heating

When stored for a long duration, it becomes darker in color. It loses some of its properties and
may also ferment if the water content is too high. Therefore, prolonged storage should be
avoided, while newly harvested honey should be preferred (Ashenafi, 2006). Heating honey
leads to drastic changes in its chemical composition. As a result, heating at high temperatures
reduces its benefits. No wonder many people prefer raw, organic or raw organic honey. While
raw by definition signifies less processing (and no heating), Organic honey is prepared using
stringent organic production methods and processing standards, in which heating at high
temperatures is not allowed (Coultate, 2009).

2.5.4. Agro-chemicals

A number of factors linked to industrial modes of agriculture affect bee colonies and other
pollinators around the world, ranging from habitat degradation due to monocultures with
consequent declines in flowering plants and the use of damaging insecticides. Incentives should
be offered to farmers to restore pollinator-friendly habitats, including flower provisioning within
or around crop fields and elimination of use of insecticides by adopting agro ecological
production methods. Conventional farmers should be extremely cautious in the choice, timing,
and application of insecticides and other chemicals (Nicholls and Altieri, 2013).

Pesticides are the chemicals that are most widely used to control pests incrop production. When
different chemicals are applied to the crops, they are affecting the pests of the crops but also
harm the beneficial insects as pollinators, predators and parasites. This harmful effect disturbs
the natural balance between the insects and their natural hosts (Wilson and Tisdell, 2001). Older
worker bees forage outside the hive for pollen and nectar, and thus are vulnerable to contact
exposure to pesticides during foraging as well as dietary exposure during collection or ingestion
of pollen and nectar. Workers also serve as a vector for bringing contaminants back to the hive.
Young workers clean cells and attend brood, whereas middle aged workers do a variety of tasks
mainly within the hive. All the young and middle-aged workers, queen and drone can have
18
secondary exposure to pesticides through contaminated food brought back to the hive (Van der
Sluijs et al., 2013).

2.5.5. Honey Bee Disease

The biology and health of the honey bee Apis mellifera has been of interest to human societies
for centuries. The bees and their products are vulnerable to various diseases, parasites and pests.
Honey bees diseases, pests and predators are causing a significant economic loss in honey bees
and their products. The most commonly known honeybee diseases reported to exist in Ethiopia
are Nosema, Amoeba and Chalk brood diseases (Evans and Schwarz, 2011). Nosema is caused
by Nosema apis and Nosema ceranae. It is a microsporidian fungal disease that infects the
intestinal tract of adult bees. Nosema cause detrimental effects on honey bees, colony
development, queen performance and honey production. In Ethiopia Nosema was reported in low
infestation rate in the survey conducted by the initiation of FAO (Fikru, 2015). In Ethiopia
Nosema was also reported from different regions with varying prevalence rate such as 58% in
Oromia, 60% Benishangul-Gumuz and 47% in Amhara regions (Teferi, 2018). Amoeba is
diseases of honey bee caused by a single celled parasite called malpighamoeba mellificae. The
parasite affects malpigian tubules of honey bees andshortens the life cycle of bees (Evans and
Schwarz, 2011).

Diagnosis made on honey bees in field and laboratory at Addis Abeba reported a prevalence rate
of 73% of amoeba infestation (Begna and Kebede, 2005). The diseases was also reported with
high prevalence rate in different regional state of Ethiopia such as; Oromia region with
prevalence rate (88%), Amhara region (95%) and 60 % in Benishangul- Gumuz (Diribe et al.,
2012). American foulbrood (AFB) is an infectious disease of the larval stage of the honeybee
Apis mellifera. It is caused by a Gram positive bacterium called Paenibacillus (Genersch, 2010).
European foulbrood (EFB) is a severe bacterial brood disease caused by the Gram-positive
bacteriumMelissocccus plutonius. The disease has a worldwide distribution and is an increasing
problem in some areas. Although the causative agent of EFB was described almost a century
ago, many basic aspects of its pathogenesis are still unknown (Forsgren, 2010).

19
2.5.6. Honey bee predators and pests

Honey bees are active defenders of their colony. Their stinging and biting behavior is very
effective at repelling intruders, even intruders as large as bears. Guard bees initiate colony-level
responses by identifying threats and recruiting nest mates for collective defense. Recruitment is
achieved through chemical communication. Volatile chemicals (pheromones) released by guards
„sound the alarm‟ and attract recruits to the entrance of the colony. Interestingly, young bees
normally stay within the hive and do not participate in colony defense (Breed et al., 2004).
However, detection and subsequent avoidance of cryptic predators, including crab spiders that
can change color to match their background pose intriguing challenges with respect to the
relative investments into decision speed and accuracy, as well as the minimization of false-
negative responses, in which over cautiousness might compromise the range of available
foraging options (Ings and Chittka, 2008).

Lice are known to infect honey bees in hive. Bee louse are wingless ecto-parasite fly which
causes significant damage bee colonies. Bee lice larvae feed on honey and pollen by tunneling
under the cell capping (Sarwar, 2016). In Ethiopia infestation of lice in honey bees was reported
from the western region of Shoa, Oromia regional state with overall prevalence rate of 42% with
highest prevalence rate 70.8% by Gemechis, 50% in Holeta and 17.1% in Jaldu (Gemechu et al.,
2014). And also in Tigray regional state reported an overall prevalence 4% in brood and 5.5%
adult bees (Godifey, 2015). According to Tsegaye, (2015) the small hive beetle was reported in
the Oromia regional state; 60% Jimma and 1.1% in Horo Guduru Wollega.

20
3. MATERIALS AND METHODS

3.1. Description of the Study Area

The study was conducted between November, 2019 and June, 2020 in Amuru district of Horo
Guduru Wollega zone of western Oromia, Ethiopia. Amuru district is located approximately
from 09°50'- 10°20'N latitude and 36°50'- 37°20 'E longitude in Horo Guduru Wollega Zone of
Oromia regional State of Ethiopia. The district is one of the thirteen districts in Horo Guduru
Wollega zone and is located at a distance of 72 km northwest of Shambu town, 405 km north
west of Addis Ababa capital city of Oromia regional state and Ethiopia. The district borders with
Amara regional state of Abay river in the north, Jardaga Jarte and Abe Dongoro districts in the
south, Kiramu distric in the west and again Jardaga Jarte district in the east.

Figure 2. Location map of study area

Source: GIS (2019)

21
As information obtained from the district Agriculture and Rural Development Office
(AWARDO, 2011) there are 18 rural and 3 urban kebele administrations in the district. Obora
town is the administrative center of the district. The total population of the district is estimated to
be 102,721 with the 52,398 and 50,323 male and female, respectively. From the total population
9548 (9.3 %) were urban dwellers. The majority of the inhabitants are Ethiopian protestant, with
50.81 % ,, while 15.1 % of the population are muslim, 29.86% are orthodox, and 4.23%
practiced wakefata in the district (WHO, 2013).According to AWARDO (2011), the altitude of
the district ranges from 760 - 2505 m above the sea level and the average annual rainfall and
temperature are about and 18 0C, respectively. It has three agro-ecologies where 14.29 % is
highland, 57.14 % midland and 28.57 % are lowland agro-ecology. The district is dominantly
mid altitude by agro-climatic condition (AWARDO, 2011).

The dominant and important trees in the district was lafto (Acacia species), bargamo (Eucalyptus
spp.), wadessa (Cordia Africana), kombolcha (Dovyalis abyssinica), kello (Bidens spp.) and
other trees, shrubs and climbers that provide nectar and pollen for honey bees (AWARDO,
2011). According to the district agricultural office information, the major soil types, in the
wereda include red soil, clay, black soil and brown soil. The rainfall pattern of the Wereda
generally is bimodal; the minor raining season starts in January and ends in April, while the main
rainy season begins around May and stops in September. There are about four major rivers
(Abay, Hanger, Walage and Kachalu) and many springs in the midland part of the
Wereda(AWARDO, 2011).

The livelihood of the people in the district is very diverse and the main economic activities are
mixed farming (crop farming, livestock rearing and beekeeping) is the mainstay for the majority
of the population in the area. The main types of crops cultivated by the farmers in the area are
maize, wheat, teff, nug, barley, beans, pea, coffee, chat, fruits and vegetables. The livestock
population of the district is estimated to be 245,462 in 164,898 cattle, 15,152 sheep, 20,401
goats, 7694 donkeys, 176 horses, 247 mules, 34,892 poultry and 11,431 managed honeybee
colonies. Livestock is considered as an important component in the farming system and
beekeeping also practiced mostly as income generating activity of the Wereda (AWLRDFO ,
2011).

22
3.2. Study Design

A cross-sectional studysupported by questionnaire survey and observation was conducted from

November to December, 2019 to evaluate honey physicochemical property in different agro
ecologies, beehives and beekeepers perception towards honey health benefits and beekeeping
system from the beekeepers living in the study area. The sampling units were households
keeping honey bee colony and honey retailers. Beekeepers in the three agro-ecology and honey
retailers represented the study population of the district.

3.3. Sampling Method, Sample Size Determination and Data Collection

This study had two components: the first part was laboratory analysis which was aimed at
determining the quality of honey produced in the study area. The second part was a survey which
was conducted to assess the knowledge, attitudes and practices (KAP) of beekeepers towards
honey health benefits and beekeeping systems. Prior to sample collection, cooperation letter was
sent to Horo Guduru Wollega LRDF Zonal Office from Jimma university post graduate program
for selecting beekeeping potential district and Horo Guduru Wollega Zonal LRDFO sent
cooperation letter to AWLRDO for sampling in each kebeles. Reconsensus survey and informal
data collection from the district Livestock office and key informants discussion were employed
before the actual data collection work was started.

The study district (Amuru), consists a total of eighteen rural and three urban kebeles. For this
study, the district was stratified into three agro-ecologies (highland, midland and lowland with 3,
12 and 6 kebeles, respectively), such that each stratum was made up of kebeles sharing similar
characteristics. Approximately 29 % of the kebeles were sampled from the total 21 kebeles based
on the proportion of kebeles in each agro-ecological zone. Therefore, six representative sampled
kebeles, 1 from highland, 3 from midland and 2 from lowland were selected from 18 beekeeping
practiced kebeles using purposive sampling method proportional to the beekeeping practiced
entire kebeles of the district were identified. For honey quality analysis, a total of 24 samples: of
which, 18 freshly harvested honey samples from the three agro-ecologies by considering three
hive types (modern, transitional and traditional) (1x 3) + (3 x3) + (2 x3) and another 6 samples
from the honey retailers in Obora town were collected (Table 2).From list provided beekeepers

23
in kebeles were selected for sample collection. Fresh honey samples from farm gates were
collected during night local time (6:00 - 8:00 PM) in order to minimize the biting behavior of the
bees. From a total sampling frame 1,325 beekeepers in the district 125 respondents from the
three agro-ecologies were selected for questionnaire survey by using the formula (Cochran,
1977). The individual beekeeper selection from selected kebeles was employed by using
systematic random sampling technique for interview.

3.3.1. Honey sample for honey quality analysis

3.3.1.1. Honey sample size determination and sample types

A total of 24 honey samples (0.5 kg each) which is stated by Bogdanov, 2009 (0.5 -1 kg), were
collected from purposively and conveniently selected six (6) potential beekeeping kebeles
including Sammo illamu, Mekeno, Ejere Goromti, Tombe Dangab, Chidati and Gulufa
considering their agro-ecologies and hive type they owned. Fresh honey samples were collected
during the peak honey harvesting season (November to December, 2019 locally known as
“damma tuufoo”) and retailers (mana bookaa fi daadhii) in Amuru district.Of which eighteen
(18) samples of locally produced honey (0.5 kg each) were collected from beekeepers from six
purposively and conveniently selected kebeles, were named as HHS-1 through HHS-3, MHS-4
through MHS-12 and LHS-13 through LHS-18, whereas six samples were collected from honey
retailers purposively and named RHS-19 through RHS-24 where HHS,MHS, LHS and RHS
refers to Highland Honey Sample, Midland Honey Sample, Lowland Honey Sample and
Retailers Honey Sample respectively. The number shows the sample number (Table 2).

24
Table 2. Summary of purposively selected kebeles and collected honey samples

No. Honey from hive types Agro- ecologies

retailers (RHS)
Sample from

Sample from
Highland Midland Lowland

beekeepers

Total
(HHS) (MHS) (LHS)

1. From traditional beehive 1 3 2 6

2. From transitional beehive 1 3 2 6 6

3. From modern beehive 1 3 2 6

Subtotal honey sample 3 9 6 18 6

Total honey sample collected 18 6 24

N= number of collected honey sample

3.3.1.2. Sample preparation and handling procedure

Starting from sample collection, preparing, straining, storing and handling samples, the
following sample handling directions and procedures were followed from farm-gate and retailers
house up to laboratory (QSAE, 2005). The required 0.5 kg sample for the laboratory analysis was
taken in to securely closed, chemical free, properly cleaned and dried plastic containers without
exposing to damp air, dust, dirty, light and smoke. Precautions were taken to protect the
samples, the sampling instrument and the containers from contamination by using cool jar. Each
container was sealed air-tight and marked with full detail of sampling code number and other
important particulars. Sample preparation involves identification and grouping with respect to
agro-ecologies, type of beehives, harvested date and source of collection, (Fig.3).

25
a. During harvest b. Straining and preparing of honey

Figure 3. Prepared and labeled honey samples for laboratory analysis

3.3.1.3. Laboratory methods and procedures

The collected honey sample were prepared according to the “COMESA 002 (2004) Standard for
Honey” protocol for the quality analysis.The quality parameters of the interest were: moisture
content (%), HMF (mg/kg), pH, free acidity (meq/kg), ash content (%), EC (mScm-), reducing

26
sugars, (%), sucrose (%) and color (mm) of the samples were determined according to the IHC,
(Bogdanov, 2009). There is quality standards of the honeywhich is stated by national and
international norms (see table 4 below). All parameters were measured according to the
internationally harmonized commission methods of the honey commission of (2009). The result
of the laboratory analysis of honey samples was compared with Quality and Standards Authority
of Ethiopia (QSAE, 2005), Codex Alimentarius Commission (CAC) and European Union
(EU).The physicochemical properties of the collected honey samples were analyzed at Holeta
Bee Research Center (HBRC) analysis laboratory. The procedure, principle, reagents,
equipment‟s used and procedure followed to do both physical and chemical analysis of honey
sample according to (IHC, 2009) was elaborated in (Annex 3 to 11).

3.3.2. Questionnaires

Semi- structured questionnaire was prepared both close and open-ended questions were included
(Annex 15). Verbal consent was obtained and the objectives of the study were explained for the
respondents. Questionnaire was intended to the owners of beehives/ beekeepers to get
information related to socio-demographic characteristics, purpose of beekeeping, beekeeping
system, honey bee flora, beekeeping constraints and KAP of beekeepers towards health benefits
of honey through face to face conversion. Beekeeping system, type of beehives, management and
KAP commonly practiced in the study area was collected from district LFRDO, DA`s record and
personal communications of selected beekeepers. The questionnaire was pre-tested and adjusted
as required translated in to local language (Afaan Oromo) for interviewees. Information collected
was ethically respected and beekeepers interviewed from kebeles were proportionally selected
from each agro-ecology.

Determining sample size in each agro-ecology and kebele was proportional to entire population
of households. Therefore, From a total of six representative kebeles namely; Samo Ilamu from
highland, Mekeno, Ejere Goromti and Tombe Dangab from midland, Chidati and Gulufa from
lowland were selected purposively based on having large number of beekeepers, beekeeper
experience, and potential area for beekeeping, abundance of honey bee colony, availability of
common bee flora and agro-ecology of the district.The sampling units were households keeping
honey bees. The sample size required for the study questionnaire survey was determined by the
27
formula recommended by Cochran (1977) and Pandey and Verma, (2008) determination
formula.

Accordingly, a total of 125 respondents‟ were selected for the interview from the district as
follows. 5% sampling error was used as a standard.
no = Z2 p q / e2 ……………………………………………………(Cochran, 1977)

= 138.29 after this by using finite population correction factor

n1= 138.29 / 1 + [ 138.29/1325｝

= 125beekeepers were selected from the district for interview

Where;
no= desired sample size Cochran‟s (1977) when population greater than 10,000
n1= finite population correction factors (Cochran‟s formula, 1977) less than 10, 000
Z = standard normal deviation (1.96 for 95% confidence level)
P = 0.1 (proportion of population to be included in sample i.e. 10%)
q =is 1-P i.e. (0.9)
N = is total number of population
d =is degree of accuracy desired (0.05)
After determining the total sample size from all agro-ecologies of the district to select
interviewers from each agro-ecology the following formula was used.

n * N1 n* N2
n1  and n2  (Pandey and Verma, 2008)
N N ……………….

Where;

n1, n2 and n3 = is sample size of respondent in each agro ecology

So from this:
n1= 125x134/ 1325= n1= 13 respondents were selected from highland agro-ecology
n2= 125 x 673/1325 = n2= 63 respondents were selected from midland agro-ecology

28
 n3= 125x518/1325 = n3= 49 respondents were selected from lowland agro-ecology
N1 and N2 = is total number of household in each agro ecology
n= total sample size of respondent in three agro ecology i.e. 125
N = is total number of beekeepers in the study area
After determining sample size from each agro-ecologies, we have selecting each beekeepers by
usingsystematic random sampling method.

Table 3. Total selected kebeles and beekeepers for questionnaire survey

Kebeles / Beekeepers/Beehives Amuru district

High land Mid land Low land Total

All kebeles found in the district 3 12 6 21

Kebeles where beekeeping is practiced 3 9 6 18
All beekeepers of the district 134 673 518 1,325

Selected respondents for the study 13 63 49 125

3.4. Data Analysis

Data was managed both in hard and software. The evaluation of honey quality was subjected to
statistical analysis using General Linear Model (GLM) of SPSS version.23. In all the analyses, P
<0.05 is set for significance and not significant as P > 0.05 level was used to separate the means
whenever GLM showed statistically significant difference. Collected data from both primary and
secondary sources was documented, organized, analyzed and summarized using MS excel and
SPSS version 23 for descriptive statistics. The survey data was coded and tabulated for analysis
using SPSS statistical package version. Simple descriptive statistics was employed in order to
have a summary description of the data collected from the survey. This involves the use of
percentages, means, frequency distributions and standard deviations to describe parameters such
as socio-demographic characteristics, KAP of beekeepers, beekeeping costraints and so on.

29
4. RESULTS

4.1. Results of Honey Quality Parameters

In this finding the results of honey quality analysis of the total of 24 honey samples, of this 18
obtained from beekeepers and 6 from honey retailers of Amuru district in the western part of
Ethiopia are presented, in(Table 4, 5, 6, 7 and 8 below). Honey samples acquired purposively
from 6 kebeles considering their agro-ecologies and beehives and 6 from honey retailers, were
subjected to analyses with regard to moisture, HMF, pH, free acidity, electrical conductivity, ash,
reducing sugar, apparent sucrose, color and their results are well summarized. In the present
study measurements were performed twice and the average result was taken and most of the
overall mean of quality parameters evaluated were indicated in agreement with national and
international standards (Table 4 and 5).

30
Table 4. All mean results of honey samples collected from the study area

Overall Sources @ Standards

Maximum

Minimum
Mean ± SD Mean ± SD (N=24)

Range
Quality Beekeepers Retailers
Parameters N (n=18) (n=6) National International
MC (%) 24 20.43± 1.32 22.8 18 4.8 19.90± 1.04* 21.90± 0.79 * 17.5-21 18-23
HMF(mg/kg 24 16.39 ± 2.68 39.2 0 39.2 10.76 ± 9.47* 33.28 ± 5.78* < 40 40-80
pH 24 3.92 ± .07 4.8 3 1.8 4.05 ± 0.24 * 3.54 ± 0.39 * - 3.2- 4.5
FA (meq/kg) 24 36.67 ± 2.13 55 14 41 33.00 ± 9.23* 47.67 ± 3.98 * < 40 5-54
Ash (%) 24 0.25 ± 0.14 .50 0 .50 0.26 ± 0.15 0.22 ± 0.12 0.6 0.25 – 1
EC (mS/cm) 24 0.68 ± 0.3 1 0.1 0.9 0.6 ± 0.3 0.5 ± 0.2 < 0.6 0.22-1.52
RS (%) 24 73.08 ± 0.92 80 66 14 73.22 ± 4.37 72.67 ± 5.32 65 60-70
AS (%) 24 1.80 ± 0.35 0 4.8 4.8 1.72 ± 1.70 2.07 ± 1.91 10 3- 10
Color (mm) 24 103.75± 2.89 129 75 54 98.94± 12.45* 118.17± 8.08* - -

@
Quality and Standards Authority of Ethiopia (2005)
*= Means within a row and column are significantly different (P < 0.05) both in univariate and multivariate GLM analysis
N= total number of sample, SD= Standard deviation MC =moisture content, HMF = Hydroxyl Methyl Furfural, FA = Free Acidity,
Ash = Total ash content, RS = Reducing Sugar, AS = Apparent Sucrose, EC = Electrical conductivity)

31
Table 5. Mean results of honey samples collected from different Agro-ecology and hive types

Agro-ecology Hive types

(Mean ± SD)(n=18) (Mean ± SD)(n=18)
Quality Highland Midland Lowland Modern Transitional Traditional
Parameters (n=3) (n=9) (n=6) (n=6) (n=6) (n=6)
Moisture (% ) 20.50± 1.21* 20.50±.52* 18.80±0.53* 20.30±0.82 20.10±1.01 19.90±1.09
HMF (mg/kg) 1.62±.62* 6.37±1.08* 22.15±2.65* 6.08±2.71 11.85±4.24 14.35±4.28

pH unit 4.02±0.06 3.94±0.03 4.22±0.14 4.20±0.13 3.99±4.11 3.95±0.07

FA(meq/kg) 28.33±0.28* 38.44±2.02* 27.17±4.30* 26.50±3.22 35.50±2.93 37.00±4.05

Ash (%) .13±0.06 .29±.18 .28±.13 .22±.01 .20±.18 .26±.20

EC (mS/cm) .40±.1 .6±.3 .7±.2 .5 ±.2 .5 ±.3 .6 ±0.3

RS (%) 72±1.53 73.78±1.29 73±2.48 71.83±1.70 75±1.51 72.83±2.15

AS (%) 0.83±0.38 1.26±0.58 2.85±0.64 2.18±0.84 1.93±0.66 1.03±0.59

Color(mm) 103.67±3.79 99.78±13.82 95.33±13.62 101.5±11.33 100.27±3.79 98.25±13.0

* = Means in the row and column is means which revealed significant difference at (p < 0.05)

4.1.1. Moisture Content

The overall mean value of moisture for the honey samples acquired from the study area was
20.43 ± 1.32 %; with the lowest moisture content of 18 % obtained for the sample acquired from
beekeepers and the highest moisture content of 22.80 % obtained for the sample acquired from
honey retailer (Table 4, 5 and Annex 12). The average value of moisture content of fresh honey
sampled from beekeepers were (19.90 %) which is numerically lower than the overall mean of
honey samples obtained from retailers (21.90 %). Therefore, the moisture content of the sampled
honey for this study revealed that significant difference (P < 0.05) between honeys sampled from
beekeepers and retailers, (Table 6). In this finding the mean value of moisture content of the
honey sample collected from highland and midland agro-ecologies were equal (20.50 %); while
32
the moisture content of lowland agro-ecology is 18.80 %. From this result moisture content of
honey sampled from different agro-ecologies revealed significant difference (p < 0.05) among
agro-ecologies. The mean moisture content of the honey sampled from different hive types were
numerically 20.30 %, 20.10 % and 19.90 % from modern, transitional and traditional hives,
respectively. Even if the numerical mean value of moisture content is varied among hive types;
statistically, there is no significant different (p > 0.05) among beehives, (Annex 13). Generally,
except some value of moisture content of honey samples acquired from retailers, all honey
samples collected from the study area has moisture content of honey in the range of national and
national standards, (Annex 12).

4.1.2. Hydroxymethyl furfural (HMF)

The HMF overall average for the honeys used in this study was 16.39 ± 2.68 mg/kg; with the
lowest HMF amount of 0.0 mg/kg obtained for the honey sample from beekeepers; whereas the
highest HMF amount of 39.2 mg/kg was determined for the honey sample obtained from
retailers, (Table 4, 5 and Annex 12). The average result of honey obtained from beekeeper (10.76
mg/kg) is much lower than that of retailers (33.28 mg/kg) which indicates the freshness of honey
of beekeepers than retailers in the present study. Similarly there is significant difference (p <
0.05) between sources of honey collected from beekeepers and retailers of the study area. The
HMF mean value of the honey collected from lowland agro-ecology (22.15 mg/kg) is higher than
that of midland (6.37 mg/kg) and highland (1.62 mg/kg). Therefore, there is a significant
difference (P < 0.05) in HMF value among agro-ecologies. The honey sampled from different
hives indicated different mean values of HMF numerically 6.08 mg/kg, 11.85 mg/kg and 14.35
mg/kg collected from modern, transitional and traditional hive types respectively. Despite,
numerical difference of HMF value in different hive types there is no significant difference (P >
0.05) among hive types of this finding, (Annex 12 and 13).

4.1.3. pH

The overall mean of pH for the honeys used in this study was determined as 3.92 ± 0.07 which
ranges from 3.00 to 4.80. With the lowest pH determined for the honey sample acquired from
retailers with 3.00; whereas the highest pH was obtained for the sample from beekeepers 4.80
(Table 4). In this finding the pH numerical values of honey samples collected from beekeeper is
33
higher than honey samples obtained from retailers, So, significant difference (P < 0.05) were
observed between the sources (Table 5 and 6). The pH values of honey samples analyzed in this
study ranged from 3.00 to 4.80 with the mean value of 4.02 from highland honey, 3.94 from
midland and 4.22 from lowland agro-ecologies. The pH value of honey sampled from lowland
agro-ecology is higher than that of highland and midland agro-ecology. In other case, the overall
mean values of pH honey sampled from modern, transitional and traditional hives were 4.20,
3.99 and 3.95 respectively (Table 5). So, thecurrent finding of pH value of honey sampled was
revealed that there is no significant difference among agro-ecologies and hive types analyzed (
Table 5 & Annex 13).

4.1.4. Free acidity

The overall mean of the free acidity of the study area was 36.67 meq kg−1 with the range of 14
to 55 meq kg−1, with the lowest free acidity value obtained for the fresh honey sample acquired
from beekeepers with 14 meq/kg; whereas the highest free acidity value was obtained for the
honey sample acquired from the retailer with 55 meq/kg. There is high range (41 meq/kg)
between fresh honey sampled from beekeepers and retailers. There was a statistically significant
difference between the free acidity values of the honey samples used in this study (p < 0.05)
between beekeepers and retailers honey (Table 6 and Annex 12). The mean and range of the free
acidity of the honey sample obtained from different agro-ecologies were 28.33 meq/kg (22
meq/kg to 30 meq/kg) from highland, 38.44 meq/kg (31 meq/kg to 48 meq/kg) from midland
and 27.17 meq/kg (14 meq/kg to 46 meq/kg) from low land agro-ecology. The result of this
study was observed that there was significant variation (P < 0.05) in free acidity among agro-
ecologies. The mean of different hive types of this finding were 26.50 meq/kg from modern,
35.50 meq/kg from transitional and 37.00 meq/kg from traditional hives which is different
numerically, however, it showed no significant difference (p > 0.05) among hive types.

4.1.5. Ash content

The overall average of Ash amount for the honey samples used in the present study was 0.25 ±
0.03 %; with the lowest and highest ash amount obtained for the honey samples acquired from
beekeepers with 0.0 % and 0.50 % respectively. Therefore, there was no statistically significant
difference between the ash amounts for the honey samples used in the study (p > 0.05) between
34
of sources of honey (Table 6).The mean and range of Ash obtained from different agro-ecologies
were 0.13 % (0.1 % to 0.2 %) from highland, 0.29 % (0 % to 0.5 %) from midland and 0.28 %
(0.1 % to 0.5 %) honey sampled from lowland agro-ecologies. The ash mean value obtained
from modern, transitional and traditional hives were 0.22 % (0.1 % to 0.3 %), 0.20 (0 % to 0.5%)
and 0.37 % (0.1 % to 0.5 %), respectively (Table 5). There was no significant difference (p >
0.05) both due to agro-ecologies and hive types analyzed, (Annex 13).

4.1.6. Electrical conductivity

The electrical conductivity overall mean value of the samples used in this study was determined
as 0.68± 0.3 mS/cm (Table 4); with the lowest value obtained for the honey sample acquired
from beekeepers with 0.1 mS/cm and the highest value obtained for the honey sample acquired
both from beekeepers and retailers with 1 mS/ cm. The overall average electrical conductivity of
honey sampled from beekeepers (0.6 mS/cm) where numerically higher than that of retailers
honey sample (0.5 mS/cm) but, there was no statistically significant difference (p > 0.05)
between honey samples of beekeepers and retailers (Table 5 & 6). The mean conductivity of
honey sampled for the present study was 0.4 mS/cm (ranges from 0.3 mS/cm to 0.5 mS/cm for
highland, 0.6 mS/cm (ranges from 0.1 mS/cm to 1 mS/cm) for midland and 0.7 mS/cm (ranges
0.3 mS/cm to 1.0 mS/cm) honey sampled from lowland agro-ecology. The overall mean value of
electrical conductivity from modern, transitional and traditional hive types were 0.5 mS/cm (0.3
mS/cm to 0.7 mS/cm), 0.5 mS/cm (0.1 mS/cm to 1.0 mS/cm) and 0.6 mS/cm (0.3 mS/cm to 1.0
mS/cm), respectively, (Table 6 and 7). The value of electrical conductivity not varied
significantly (p > 0.05) due to agro-ecologies and type of beehives analyzed (Annex 13).

4.1.7. Reducing sugar

From the total of 24 honey sampled for the study the overall mean of reducing sugar (fructose
and glucose) of Amuru district was 73.08 ± 0.92 %; with the lowest and highest reducing sugar
ratio obtained for the fresh honey sample acquired from the beekeepers with 66 % and 80 %.
Numerically the highest and lowest value was obtained from beekeepers and there was a no
statistically significant difference (p > 0.05) between the total reducing sugars of the honey
samples studied of the honey obtained from beekeepers and retailers (Annex 13). The mean
reducing sugar of honey sampled from different agro-ecology and hive types were 72 % (69 % to
35
74 %) from highland, 73.78 % (71% to 80 %) from midland and 73 % (66 % to 80 %) from
lowland agro-ecologies. While 71.83 % (66 % to 78 %) from modern, 75 % (71 % to 80 %) from
transitional and 72.83 % (68 % to 80 %) obtained from traditional hive types, (Table 5). The
mean reducing sugar of honey sampled from different agro-ecologies and hive types were no
significant difference (p > 0.05) observed both due to agro-ecology and hive types, (Annex 13).

4.1.8. Apparent Sucrose

In this study the total average of apparent sucrose for the honey samples obtained from the study
area was 1.80 ± 0.35%; with the lowest sucrose calculated for the collected sample from
beekeepers with 0.00 % value; while the highest apparent sucrose ratio was obtained for the
honey sample acquired from the retailers with 4.8 % value (Table 4 and Annex 12). Despite the
mean is varied numerically, there was no statistical significant difference (p > 0.05) between
honey sampled from beekeepers and retailers. The mean sucrose content of honey sampled from
the highland, midland and lowland agro-ecologies were 0.83 % (0.1 % to 1.4 %), 1.26 % (0 % to
4.4 %) and 2.85 % (0.1 % to 4.5 %). Whereas the mean sucrose of honey sampled from modern,
transitional and traditional hives were 2.18 % (0 % to 4.5 %), 1.93 % (0.1 % to 4.4 %), and 1.03
% (0.1 % to 3.1 5), respectively, (Table 5). There was no significant difference (p > 0.05)
observed both due to agro-ecology and hive types in apparent sucrose value of the present study
(Annex 13).

4.1.9. Color

The overall average value of color of honey samples used in the study area was 103.75± 2.89
mm which indicates amber color, with this the lowest color value obtained from the honey
sample acquired from the beekeepers with 75 mm which revealed light amber color; while the
highest color value was determined from retailers honey sample with 125 mm with dark amber
color. The average color value of honey sampled of this study from beekeepers were 98.94 mm
which indicates amber color, while the color mean value of honey sampled from retailers were
117.17 mm reveals dark amber color (Table 4). From the total sample collected 62.5 %, 16.67 %
and 20.83 % were amber, light amber and dark amber color respectively. 83.33 % studied honey
sample of dark amber color was calculated from retailers honey sample (Annex 12). There was
statistically significant difference (p < 0.05) between the color values of the honey samples
36
examined from beekeepers and retailers (Table 6). The average color value of honey sampled
from different agro-ecologies were 103.67 mm from highland, 99.78mm from midland and
95.33mm from lowland which all indicated amber color, while the color mean value of honey
sampled from different hive types were 101.5 mm from modern, 100.27 mm from transitional
and 98.25 mm from traditional hive types which all mean values in the range of amber color
(Table 5). Therefore there was no significant difference (p > 0.05) both due to agro-ecologies
and hive types of honey sampled, (Annex 13).

4.2. Results of Univariate General Linear Model Analysis of Honey Quality Parameters

In this finding according to univariate analysis of General linear model (at 95 % CI and p < 0.05
significant difference) the sampled honey from different agro-ecology significantly affects the
parameters (P < 0.05) among the samples for moisture content, HMF and free acidity. However,
no significant differences (P > 0.05) in pH, Ec, ash content, reducing sugar, apparent sucrose and
color were revealed among the honey samples collected from different agro-ecologies. Moisture
content, HMF, pH, free acidity, and color were revealed significantly different (p < 0.05):
whereas, Ash content, Ec, reducing sugar and apparent sucrose revealed not-significant
difference (p > 0.05) observed between sources of honey ( i.e. beekeepers and retailers). Finally,
all parameters evaluated in this finding were not showed any significant difference (p > 0.05)
among samples collected from different hives types (Annex 13).

4.3. Results of Multivarite General Linear Model Analysis of Honey Quality Parameters

In these study variables with a p ≤ 0.25 in the univariate GLM analysis under different
independent variables like source, agro-ecology and type of beehives were included in the final
multivariate GLM analysis. Accordingly; moisture, HMF, pH, free acidity and color, were
analyzed by multivariate analysis for sources and revealed that all those analyzed parameters
have significant difference (p < 0.05)between sources (beekeepers and retailers) of honey sample
collected like in univariate analysis ( See Table 6).

37
Table 6. Multivariable analysis between sources

Independent variable Dependent No. of F P-value

variable tested
MC (%) 24 18.482 .000

HMF (mg/Kg) 24 29.699 .000

Source
pH 24 14.882 .001
FA (meq/Kg) 24 13.943 .001
Color (mm) 24 12.351 .002

In the same way; moisture, HMF, pH, free acidity and apparent sucrose were analyzed for agro-
ecology. However, only moisture, HMF and free acidity have significant difference (P< 0.05)
among agro-ecologies in multivariate analysis (Table 7). Parameters analyzed by multivariate for
hive types having p <0.25 like pH, free acidity ash content and EC value, indicated that they
couldn‟t show any significant difference (p > 0.05) among beehives. In general, in the present
study sources of honey has a significant (p < 0.05) effect on the values of honey moisture, HMF,
pH, free acidity and color and Agro-ecologies have significant (p < 0.05) effect on moisture,
HMF and free acidity value of the present analyzed honey.Whereas, type of hive have`t any
significant (p < 0.05) effect on the parameters of honey analyzed in the study area ( See Table 7).

Table 7. Multivarite analysis among agro-ecologies

Independent variable Dependent variable No . of F P-value

tested
MC (%) 18 13.939 .000
Agro-ecology HMF (mg/Kg) 18 31.116 .000
FA (meq/Kg) 18 4.412 .031

38
4.4. Correlation between the Quality Parameters of Honey Sampled

In this finding the physicochemical properties of honey of the study area had different correlation
results between each other. Pearson correlation coefficients between all parameters were
presented (Table 8). In correlation criterion there is strong negative/ positive and weak negative/
positive ranges from -1 to +1. There were strong positive significant (p < 0.01) correlations
between moisture content and free acidity and color r = 0.63 and 0.53 respectively. HMF is
significantly correlated with free acidity and electrical conductivity with the value of r = 0.45*
and 0.77**, (p < 0.05) and (p < 0.01) respectively. Whereas the overall free acidity of the honey
sample was significantly correlated with electrical conductivity with the r = 0.41 value (p <
0.05). Other parameters had shown numerical strong and weak correlation between variables.
Also in the current evaluation of correlation there is strong relationship between HMF and
electrical conductivity with the highest r= 0.77 ** (P < .01). The high correlation coefficient (r =
0.77 **) value indicated pure/ fresh honey is characterized by a low HMF and conductance value
in , (Table 8 below).

Table 8. Pearson correlation coefficients among the analyzed parameters

Parameters MC HMF pH FA Ash EC RS AS Color

Moisture (%) 1 .19 -.58** .63** -.16 .11 -.12 -.24 .53**
HMF(mg/kg 1 -.50* .45* .09 .77** .13 .14 .33
pH 1 -.79** -.05 -.32 -.29 .26 -.29
*
FA (meq/kg) 1 .00 .41 .04 -.07 .19
Ash (%) 1 .35 .11 -.14 -.09
EC (mScm-) 1 -.03 .27 .15
RS (%) 1 -.54** .13
AS (%) 1 -.34
**. Correlation is significant at the 0.01 level (2-tailed),
*. Correlation is significant at the 0.05 level (2-tailed). r= correlation

39
4.5. Questionnaire Survey

4.5.1. Socio-demographic characteristics and beekeeping system of the respondents

In this section, the major socio-demographic characteristics of households interviewed in

the survey were described. These characteristics related to the relative frequency distribution of
household heads by educational background, gender, age, beekeeping experience and livestock
they owned.From the total 125 selected respondents: 10.40 %, 50.40 % and 39.20 % of them
were selected from highland, midland and lowland agro ecology respectively, proportional to
entire beekeepers. Majority of the respondents are (about 67.20 %) under educated (below high
school) (see table 9). Of the selected beekeepers interviewed male to female ratio were of 6.82: 1
(87.20 %) male.In mid and low land there are few female beekeepers than males whereas in
highland none of them do not participating in this activity. Females active participation was
limited in the study area their support of men in collaboration with the family members in terms
of cleaning the apiary, controlling pests and predators, selling hive products, preparation and
offering supplementary feed was not ignorable.To maximize women‟s participation in beekeepin
g training should have to be offered to empower them.

The mean age distribution of the respondents were 43.31 ± 19.75 (with the range of 14 to 81
years). In this study people in the range of active and productive age groups are actively
participated in beekeeping activities. The beekeepers had an average experience 7.66 (ranges
from 3 to 26 years.) of beekeeping in the district. Of 125 respondents the livestock rearing
household heads in the study area were presented. Of the total households interviewed, were
97.60 % were cattle owners while 44 % sheep, 48.80 % goats, 72.80 % equines and 72 % rearing
poultry (Table 9).

40
Table 9. Socio-demographic indicators and beekeeping system of respondents (N= 125)

Descriptions Agro-ecologies
Highland Midland Lowland Total
(n=13) (n=63) (n=49) (N= 125)
N (%) N (%) N (%) N (%)
1. Educational Background
No informal and 2 (15.38) 6 (9.52) 11 (22.45) 19 (15.20 )
formal education
Elementary school 7 (53.85) 31 (41.27) 27 (46.94) 65 (52)
High school 3 (23.08) 20 (31.75) 9 (18.37) 32 (25.60 )
Colleges / University 1 (7.69) 6 (9.52) 2 (4.08) 9 (7.20 )
2. Gender
Male 13 (100) 51 (80.95) 45 (91.84) 109 (87.20 )
Female - 12 (19.05) 4 (8.16) 16 (12.80 )
3. Age
14-24 years 4 (30.77) 18 (28.57) 13 (26.53) 35 (28)
25-65 years 6 (46.15) 32 (50.79) 29 (59.18) 67 (53.60 )
> 65 years 3 (23.08) 13 (20.63) 7 (14.29) 23 (18.40)
4. Beekeeping Experience
< 5 years 4 (30.77) 16 (33.33) 11 (22.45) 31 (24.80 )
≥ 5-10 years 7 (53.85) 35 (55.56) 33 (67.35) 75 (60)
≥ 10 years 2 (15.38) 12 (19.05) 5 (10.20) 19 (15.20)
5. Livestock Ownership
Cattle 13 (100) 60 (95.24) 49 (100) 122 (97.60)
Sheep 5 (38.46) 24 (38.10) 26 (53.06) 55 (44 )
Goat 4 (30.77) 20 (31.75) 37 (75.51) 61(48.80 )
Equines 9 (69.23) 42 (66.67) 40 (81.63) 91 (72.80)
Poultry 6 (46.15) 51 (80.95) 33 (67.35) 90 (72)

N= Total number of respondents in study area n= number of respondents in each agro ecology,
%= percentage

In the sampled population 52 %, 33.60 % and 96.80 % of beekeepers owned modern, transitional
and traditional hive types. The average of beekeepers who owned improved beehives was 21.67
households, 14 households owned transitional hive and for beekeepers that had traditional
beehives was 40.33 households in each agro-ecology. The user of modern hive was 65.08 %

41
from midland, 46.15 % from highland and 36.37 % from lowland agro-ecologies this is may be
due to proximity of midland agro-ecologies of the interviewed beekeepers to the town of the
district and closed to get technology assess and information from the livestock office. All
beekeepers lived in the study area in highland and lowland agro-ecologies had traditional type of
hives. Transitional hive usage is mostly seen in lowland agro-ecology with 46.94 % than that of
highland and midland.

Table 10. Hive types owned by beekeepers of respondents among agro-ecologies

Hive type Agro-ecologies Total (N =125)

Highland (n=13) Midland (n= 63) Lowland (n= 49) N (%)

N (%) N (%) N (%)

Modern 6 (46.15) 41 (65.08 ) 18 (36.73) 65 (52)

Transitional 5 (38.46) 14 (22.22) 23 (46.94) 42 (33.60)

Traditional 13 (100) 59 (93.65) 49 (100) 121 (96.80)

N= total no. of respondents n=number of beekeepers in each agro-ecology

4.5.2. Knowledge, attitude and practices of beekeepers on public health benefits of honey

Respondents main purpose of keeping honey bees were 88 % for both household consumption
and income generation, 4 % only for income generation and 8 % only for household
consumption.The beekeepers have developed indigenous knowledge of keeping bees passing
from generation to generation. These indigenous beekeeping knowledge are hive construction
from locally available materials, swarm catching, hive fumigation materials, honey and
swarming season identification, different medicinal values of honey, identification of important
honeybee floras, the medicinal value of honey and identification of adulterated honey.

In these survey beekeepers experiences in purpose of using honey activities the deep indigenous
knowledge of beekeepers which may contributes to the development of modern medicine varied
42
from area to area and from agro-ecology to agro-ecology. In this finding most of the selected
respondents in the district (89.60 %) had knowledge, attitude and practices of honey health
benefits as traditionally using for many diseases, symptoms and values and (10.40 %) of the
respondents consume honey as foods without knowing the benefit of honey on the health
perspective. About 32.80 % the respondents belief that honey has side effects on infants less
than one year old but, they can`t considered as disease rather than the honey side
effect.Interviewed beekeepers replied that beekeepers had a long term experience that passed
from generation to generation (Table 11).

The knowledge of honey health benefits were mostly adopted 72 (64.29 %) from families, 34
(30.36 %) from neighbors and 6 (5.36) were experienced from other sources (book, radio,
television and etc.) Knowledge of honey health benefits has significant correlation with (p <
0.05) the beekeepers having long experience of beekeeping and age in the interviewed study
area. From the respondents interviewed the aged group has more knowledge of honey health
benefits and the highland beekeepers have 100 % knowledge of health benefits while 93.65 %
from midland and 81.63 % from lowland agro-ecologies were knew/ heard health benefits of
honey. In other hand, this research revealed that educational background; of the respondents
have no any significant (p > 0.05) relation with the knowledge, attitudes and practices of the
honey health benefits. Moreover, knowing the medicinal value of honey, identifying honeybee
pests and predators with their time of attack, method of controlling honeybee pests and predators
and cleaning hive by smoke provision from predators attack. This makes the beekeepers to know
overall things of beekeeping and essential to expand the beekeeping activity. Long term
experience as well as indigenous knowledge of beekeepers of the study district is the best
opportunity and immense potential to modernize beekeeping industry and widely utilize
available ethno-medicinal value.

43
Table 11. Knowledge, attitude and practices of beekeepers on public health benefits of honey

Descriptions Highland Midland Lowland Total

(n=13) (n=63) (n=49) (N=125)
1. Purpose of beekeeping
Both for consumption and income 13 (100 ) 51 (80.95) 46 (93.88) 110 (88 )
Only for Consumption - 9 (14.29 ) 1 (2.04) 10 (8 )
Only for income - 3 (4.76 ) 2 (4.08) 5 (4)
2. Know or heard honey public
health benefits
Yes 13 (100) 59 (93.65) 40 (81.63) 112 (89.60)
No - 4(6.35%) 9(18.37) 13(10.40)
3. From where do you get the
Knowledge
Family 9 (69.23) 38 (64.41) 25 (62.50) 72 (64.29)
Neighbor 2 (15.38) 17 (28.81) 15 (37.50) 34 (30.36)
School - - - -
Other * 2 (15.38) 4 (6.78) - 6 (5.36)
3. Know or heard on honey side
effects giving for Childs ≤ 1year

Yes 5 (38.46) 23 (36.51) 13 (26.53) 41 (32.80)

No 8 (61.54 ) 40 (63.49) 36 (73.47) 84 (67.20)

Others *= book, radio, television, story

Most of the Beekeepers interviewed in the study area were used honey alone or by mixing with
other ingredients to treat many diseases, symptoms, disorders and values by different
formulation, route of administration, frequency of using, dosage and species they treat as
traditional healer for peoples and their livestock (Table 12). Therefore, in this finding almost all
(89.60 %) of the respondents knew the health benefits of honey traditionally. The main diseases,
symptoms, disorders and values they treat obtained

44
from questionnaires were gastrointestinal tract, respiratory, skin, reproductive and they used as
cosmotics specially by females. The scientific and local names of disease and other values were
stated in the (Table 12). The formulation of the honey for medicinal value is bolus, syrup, juice,
powder and infusion forms. The routes of administrations stated by the respondents were 54.55
% oral and 45.45 topical. The frequency and dosage of drugs given is mostly as needed, from 1,
2 and 3 days as administered by the beekeepers local healers. The species treated by the honey
and additives were human being and livestock, (Table 12).

45
 Table 12. Summary of medicinal value of honey in the study area

Indication Disease local Form of preparation Route Frequency Dosage Spp. it

names treats
Diarrhea Garaa kaasaa Bolus: by pounding 3-4 tea spoon of honey with coffee Oral For 2-3 days morning 1-2tea spoon Human
powder and evening before meal
Abdominal Garaa dhukkubbii Juice: by mixing 1-2 tea spoon of honey with garlic Oral As needed 1-2tea spoon Human &
pain Cattle
Common cold Utaalloo Infusion: by mixing 2 tea spoon of honey with 1 raw egg Oral As needed 1cup of Human
coffee
Asthma Asmii Infusion: by mixing 3-4 tea spoon of honey with ½ lit. Oral For 2-3 weeks ½ liter Human
warm water and garlic at morning

Hangover Machii Infusion: by mixing 3-4 tea spoon of honey with 1/2lit. of Oral Stat ½ lit. Human
water

Wound Madaa Juice/ Sap: mixing 1 tea spoon of honey with salt Topical Morning& evening for 3 1tea spoon Human &
days Animals
Cuts and Burns Muraa fi Gubaa Juice: mixing 2-3 tea spoon of honey with yoghurt Topical Morning evening for 1-2 1-2 tea Human &
weeks spoon Animals
Dermatophytosi Baarollee Poultice: mixing honey with lemon, endode and salt Topical For 1-2 weeks As needed Human
s
As cosmetics Miidhaginaaf Paste/ Poultice: mixing 1 coffee cup of honey with Topical Evening before sleep as 1coffee cup Humans
papaya, yoghurt and lemon (face) needed mostly
females
Increase Fedhii saala dhiiraa By mixing honey with red teff as beverage as drinking Oral As needed From 2-3 Humans
male sexual dabaluu glass as
Potency needed

Wart Kintaarotii Mixing honey with salt powder and dissolve with cold Topical After washing with soap 1 tea spoon Human
water morning and evening of honey

46
4.5.3. Honey bee flora and their flowering season

Honeybee flora and trees of the current study area are considered to be an essential indicator for
potentialities for beekeeping activity in the district of the study area. According to the results of
this survey, the honey bee plants of the study area were trees, shrubs, herbs and cultivated crops
which are a source of nectar and pollen. Some important honey bee plants of the areas were
recorded in local and scientific names with flowering seasons from the beekeepers and
observations on the field (Table 13). The honey bee flora of Amuru district was perennial trees
and annual shrubs, cultivated crops and some herbs having significant contribution of the honey
production. This wide range of plants and vegetation variety of the area could be potentially
suitable for effective quantity and quality production of honey in various seasons. Experienced
beekeepers familiar with the plants that produce nectar or/and pollen, when it blooms and how
long they remain in blooming.

Most beekeepers of the district knew the plant flora type and medicinal value of honey depending
on the color and test of honey. The annual production of honey mostly comes from trees like
Makanisa (Croton macrostachys), Bargamo (Eucalyptus spp), Waddessa (Cordia Africana) and
Baddessa (MavlaVerticillate) usually flowers from March to June. Many beekeepers in the study
area explained that annual shrubs, crops and herbs like Hadaa (Bidens spp), Siddisa (Trifolium
Spp) and Nugi had yellow color and thick viscous honey. Respondents stated that the flowering
season of this herbs, crops and shrubs were totally from September –November, (Table 13) and
the medicinal value of this honey was very higher than autumn harvested honey from April to
May.

Figure 4. Some flowering plants in the study area

47
Beekeepers in the study area replied that they depended on many kinds of flowering plants
including trees, to be indicator shrubs, undergrowth and different kinds of flowering crops and
weeds. According to the beekeepers view, most important bee flowering shrubs, crops and weeds
flowered between September and November and most important plants flowered in March to
May and some in January. In Amuru district, the major honey harvesting season is between
October and December from Gizotia scabira, Bidens pachyloma,Cordia Africana, crops (maize.
wheat and barley) and other weedy species. Local names, and flowering seasons are given based
on field observations and local knowledge of beekeepers (Table 13).

Table 13. Major honey bee floras and blooming season in the study area

No. Scientific Names Local Names Blooming Season

1. Vernoia amygdalina Eebicha December-February

2. Cordia Africana Waddeessa January- July
3. Dovyalis caffra Kombolcha May –June
4. Coffea arrabica Buna April- May
5. Acacia abyssinica Laaftoo March- September
6. Ekebergia capensis Somboo September –November
7. Ficus vasta Qilxuu October- December
8. Bidens spp Hadaa/ Keelloo September –December
9. Trifolium spp Siddisa July – October
10. Zea mays Boqqolloo September- October
11. Mavla Verticillate Baddessaa April- June
12 Guizotia abyssinica Nuugii October –December

13 Schefflera abyssinica Gatamaa May – June

14 Syzygium guineese Waaleensuu October- November

4.5.4. Beekeeping challenges in the study area

Based on the results obtained from this study, beekeepers of Amuru district have faced with a
number of challenges and difficulties that affect their desired production. Major challenges in
beekeeping a rises from environmental factors that are beyond the control of the beekeepers,
while others have to do with poor marketing, infrastructure and management system. Beekeepers
were interviewed to list the constraints in order of their importance. According to the beekeepers
response and information the major challenges of the beekeeping are detailed. The major
constraints of beekeeping in the study area were pests and predators (38.40 %) (Table 14).

48
Table 14. Major constraints of beekeeping identified in the study area (N= 125)

Major constraints N (%) of respondent who experienced

constraints

Pests and predators 48 (38.4)

Agro-chemicals 26 (20.8)

Equipment and price of hive 17 (13.6)

Disease 15(12)

Deforestation 11 (8.8)

Market problem 5 (4)

Lack of beekeeping knowledge 4 (3.2)

49
5. DISCUSSION

This study is conducted in Amuru district of Horo Guduru Wollega to evaluate honey quality and
assess the knowledge, attitude and practices of beekeepers from November, 2019 to June,
2020.The moisture content is the most essential quality component of honey, because the rate of
fermentation, its shelf life span and processing characteristics are greatly determined by the
amount of moisture content (Gebremedhin et al,. 2013).As indicated in this finding themean
value of moisture is 20.43 %. The present study nearly agrees with findings of Belay et al. (2013)
who reported 20.50 % as mean test result for Ethiopian honey. However, the finding islower than
the average honey sample (22.86 %) reported by Getachew et al, (2014).This finding much
higher than the average moisture content of (18.80 %), (17.89 %) and (14.41 %), of honey
moisture reported by Belay et al, (2013) ; Getu and Birhan (2014) and Tesfaye et al, (2017) in
Harenna and Bale forest southeastern Ethiopia and in and around Gondar town, respectively.

Honey moisture determines the capability of honey to remain stable and resist spoilage by yeast
fermentation: the higher the moisture, the higher the probability that honey will ferment
uponstorage (Getu and Birhan, 2014).Lower moisture limits (e.g. 19%), ensuring a better shelf-
life of honey which would be met by a large majority of the commercial honeys, have been for
the revision of the Codex Alimentations (Oddo and Bogdanov, 2004).The mean value of the
moisture content of honey collected from beekeepers (19.90 %) was lower than the mean of
honey samples obtained from retailers (21.90 %).This variation might be freshness of honey from
beekeepers and hygroscopicity, poor handling and storage of honey samples obtained from
retailers (Bogdanov, 2004). The moisture content of honey depends on various factors such as the
harvesting season, the degree of maturity that honey reached in the hive, type of hive used, and
environmental temperature (Nigussie et al., 2012).

HMF: HMF is defined as a breakdown product of fructose that is formed slowly and naturally
during the storage of honey and much more quickly when honey is heated (Getachew et al.,
2014). One of the most commonly monitored parameters for determining honey freshness and
good practices by beekeepers are HMF(Pasias et al., 2018). As indicated in the HMF value of
the honey samples in this study ranges from 0 mg/kg to 39.20 mg/Kg with the mean value of
16.39 mg/kg which is within acceptable range set by QSAE, CAC and EU, i.e. 40, ≤ 40 and ≤ 60
respectively. Similarly, Tesfaye et al, (2016) have also reported that mean HMF content of
different honey samples ranged from 27.10 to 40.80 mg kg-1. In fresh honey, HMF is present
only in small amounts and its concentration increases with storage time and prolonged heating of
honey (Da Silva et al., 2016). The average result of honey obtained from beekeeper (10.76
50
mg/Kg) is much lower than that of retailers (33.28 mg/Kg) which indicates the freshness of
honey of beekeepers than retailers in the present study. Similarly, it showed significant difference
(p < 0.05) among the sources of honey. The average HMF (16.39 mg/Kg) value of the present
study was much lower than that of Adaba (38.81 mg/kg) and Dinsho (33.86 mg/kg) districts
which is evaluated by Tesfaye et al, (2016). The HMF mean value of the honey collected from
lowland agro-ecology (22.15 mg/Kg) is higher than that of midland (6.37 mg/Kg) and highland
(1.62 mg/Kg). Therefore, the finding revealed statistical significant difference (p < 0.05) among
agro-ecologies. The honey sampled from different hives indicated different mean values of HMF
numerically 6.08 mg/Kg, 11.85 mg/Kg and 14.35 mg/Kg collected from modern, transitional and
traditional hive types. Despite, numerical difference of HMF value in different hive types there is
no significant difference (P > 0.05) between hive types of this finding.

pH: Most honeys are acidic and have low pH values. The overall mean of pH for the honeys used
in the study was determined as 3.92 ± 0.07 which ranges from 3.00 to 4.80. With the lowest pH
determined for the honey sample acquired from retailers with 3.00; whereas the highest pH was
obtained for the sample from beekeepers 4.80. All honeys are acidic with a pH-value generally
lying between 3.50 and 5.50, due to the presence of organic acids that contribute to honey flavor
and stability against microbial spoilage (Bogdanov et al., 2004). All pH value of this study is in
line with pH values reported earlier. This parameter has of great significance during the
extraction and storage of honey as it influences the texture, stability and shelf life of honey
(Gomes et al., 2010).

According to Tesfaye et al, (2017), the average pH of honey samples in Dellomenna (3.92)
district of Bale Natural Forest, Southeastern Ethiopia is similar to this study. A published report
indicates that the pH of honey should be between 3.20 to 4.50 (Kinate et al., 2010). In this
finding the pH numerical values of honey samples collected from beekeeper is higher than honey
samples obtained from retailers, similarly, there is significant difference (P < 0.05) were
observed between the sources.

In the current study, the mean value of pH honey sampled from modern, transitional and
traditional hives were 4.20, 3.99 and 3.95 respectively (Table 5). So, a current finding revealed
that there is no significant difference between hive types and agro-ecologies. Similarly, the result
investigated by Gobessaet al, (2012) in Homesha district of Benishangul Gumuz was showed
that there is no significant difference (p > 0.05) in pH were observed between honey samples
obtained from different hives and also between honey samples obtained from the different
locations. In general the pH value of the present study was agreed with that of Belay et al,(2013),
51
Kinate et a,l (2013) and Alvarez et al, (2009) who were reported the pH values of 3.53 to 5.01,
3.45 to 4.18 and 3.47 to 4.24 respectively.Therefore, in the current result the low pH of honey
confirms that it well inhibits the presence and growth of micro-organisms and makes honey
compatible with many food products in terms of pH. The variations in pH might mainly be
resulted due to different acids found in different floral types. The low pH of the honey samples
showed that they are acidic and this indicated their ability to inhibit the presence and growth of
microorganisms (Ward, 2014).

Free acidity: Free acidity has been used as a quality criterion for assessing whether fermentation
has taken place; honey fermentation causes an increase in acidity. The overall mean of the free
acidity of the study area was 36.67 meq kg−1 with the range of 14 to 55 meq kg−1, with the
lowest free acidity value obtained for the fresh honey sample acquired from beekeepers with 14
meq/kg; whereas the highest free acidity value was obtained for the honey sample acquired from
the retailer with 55 meq/kg. There is high range (41 meq/kg) between fresh honey sampled from
beekeepers and retailers, which is in line with the range of national (< 40) and international (<
50) standards (Alemu et al., 2013)except honey sample obtained from retailer which is 55 meq
kg−1. There was a statistically significant difference between the free acidity values of the honey
samples used in this study (p < 0.05) between beekeepers and retailers honey.The result of this
study was in accordance with the observation made by Belay et al, (2013) ofthe mean free acid
content of the Harenna forest honey samples was 34.57 meq/kg (ranges from 25.49 to 48.81
meq/kg).

Ash content: as indicated in the above result the ash content of the honey samples obtained
ranged from 0.00-0.50g with the mean value of 0.25g. The result is consistent with (Tesfaye et
al., 2016), who reported the overall mean 0.21 % ash content of honey samples of Bale natural
forest, Southeastern Ethiopia. The ash percentage found in honey expresses its richness in
mineral content and constitutes a quality parameter.The mineral content of blossom honey ranges
from 0.1 to 0.3 % (Bogdanov, 2009). The present study revealed average ash content of honey
collected from beekeepers was numerically higher than the average ash content of retailers. In
other hand, the average ash content with agro-ecologies were 0.13% highland, 0.29 % midland
and 0.28 % lowland and also there was numerical difference between hive types as 0.22 from
modern, 0.20 from transitional and 0.37 from traditional. But, there was no significant difference
(P > 0.05) in ash content of honey samples from all agro-ecologies, type of hives and beekeepers
and retailers. This is because ash content of honey depends on the material contained in the
pollen collected by the bees during foraging on the flora.

52
In highland there were higher plant coverage than the midland and lowland. Due to this there will
be higher organic matter content in the soil. This makes the nutrient holding capacity of highland
soil to be high and higher accumulation of minerals which can be easily available to plants. This
result is also agrees with findings of Getu and Birhan (2014) in and around Gondar town and in
contrast with Gobessa et al, (2012) in Benishangul gumuz which is 0.23 % and 0.17 %
respectively. Therefore, the average result of current study revealed that honey produced in
Amuru district was in the international limit range which is 0.25-1% and which is good for
consumption and industrial purposes.

Electrical conductivity: electrical conductivity depends on ash, organic acids, proteins and some
complex sugars, and varies with botanical origin. Electrical conductivity is widely used for
discrimination between honeydew and blossom honeys and also for the characterization of
unifloral honeys (Chefrour etal., 2009).The overall mean of this study of conductivity was 0.68
mS/cm and ranges 0.2 to 1mS/cm which agrees with the international standards (0.22 to 1.52
mS/cm). The current finding of the mean electrical conductivity of the study area of 0.68± 0.018
was in line with the finding of (Getu and Birhan, 2014) who reported the mean electrical
conductivity of 0.62 Amahara region in and around Gondar. The mean electrical conductivity
reported by (Belay et. al, 2013) from Bale Harenna forest honey was slightly higher than the
current finding. From this finding is numerical difference between agro-ecologies in such that
0.33 mS/cm from highland, 0.54 mS/cm from midland and 0.78 mS/cm from lowland. And also
from this finding there is variation between beekeepers and retailers honey of the study area,
however, there is no significant difference (p > 0.05) among agro-ecologies and source of
collected honey.

The conductivity is a good criterion of the botanical origin of honey and thus is very used in
routine honey control (Oddo andBogdanov, 2004).A lower limit has been proposed for blossom
than for honey dew honeys (Pita-Calvo and V`azquez, 2017). The conductivity measurement
collected honey samples from analysis of different location with respect to Amuru district; is in
the range of 0.3 - 0.4 mS/cm, 0.2 - 1 mS/cm, and 0.7 - 0.9 mS/cm in case of highland, midland
and lowland respectively was in line with Yadata (2014) from different areas of Tepi. The
conductivity value from lowland agro-ecology is greater than in two cases of hives. According to
the current study conducted the conductivity was 95.83 % found to be less than 1mS/cm.

Reducing sugar: Sugars comprise about 95- 99% of honey dry weight and are the main
constituents of honey. Glucose and fructose are the main sugar constituents of honey and they are
produced by sucrose hydrolysis and represent 85-95% of the total sugars (Rebiai and Lanez,
53
2014). From the total of 24 honey sampled for the study the overall mean of reducing sugar
(fructose and glucose) of Amuru district was 73.08 ± 0.92 %; with the lowest and highest
reducing sugar ratio obtained for the fresh honey sample acquired from the beekeepers with 66 %
and 80 %. The general standard for a minimum content of the sum of fructose and glucose is 60
g/100 g for all blossom honeys and 45 g/100 g for all honeydew honey (Bogdanov.et al, 1999).
Accordingly, about 100 % honey samples qualify national and international standard for content
of reducing sugars in honey.

The highest and lowest value was obtained from beekeepers and there was a no statistically
significant difference (p > 0.05) between the total sugar amounts of the honey samples studied of
the honey obtained from beekeepers and retailers. The mean reducing sugar of honey sampled
from different agro-ecology and hive types were 72 % (69 % to 74 %) from highland, 73.78 %
(71% to 80 %) from midland and 73 % (66 % to 80 %) from lowland agro-ecologies, while 71.83
% (66 % to 78 %) from modern, 75 % (71 % to 80 %) from transitional and 72.83 % (68 % to 80
%) obtained from traditional hive types. In this study numerically the mean RS content of
traditional hive is higher than that of the frame hive. This is b/c all traditional hives in the area
were hanging on selected tallest tree, while the frame hives were on the ground. There is a
general truth that stated by Belay et al, (2013) during his finding in Bale as height increase the
possibility of evaporation increase. So, the traditional hive helps to concentrate the solid
component of honey which increases reducing sugar.

The mean reducing sugar of honey sampled from different sources, agro-ecology and hive types
were no significant difference (p > 0.05) observed both due to sources, agro-ecology and hive
types, (Table 6 and 7). The current finding of 73.08 ± 0.92 %; higher than the finding of
(Getachew et. al, 2014), who reported 66.79 ± 6.96 %, (Behonegn, 2017) who reported 64.3 % in
South Wollo zone and (Getu and Birhan, 2014) who reported 67.83 % in and around Gonder.

Apparent Sucrose: Analysis of sucrose content is used to detect the adulteration of honey with
table sugar or to determine the amount of sucrose naturally found in a given honey sample (Guler
et al., 2007).In this study the total average of apparent sucrose for the honey samples obtained
from the study area was 1.80 ± 0.35%; with the lowest sucrose calculated for the collected
sample from beekeepers with 0.00 % value; while the highest apparent sucrose ratio was
obtained for the honey sample acquired from the retailers with 4.8 % value. The result showed
that 100 % of the samples were in the acceptable range set by QSAE i.e. 10g/100g, the range of
CAC and EU i.e. < 5 g/100g. Despite the mean is varied numerically, there was no statistical
significant difference (p > 0.05) between honey sampled from beekeepers and retailers.
54
Higher sucrose contents could be the result of an early harvest of honey, i.e., the sucrose has not
been converted to fructose and glucose (Gomes et al., 2010). The mean sucrose content of honey
sampled from the highland, midland and lowland agro-ecologies were 0.83 % (0.1 % to 1.4 %),
1.26 % (0 % to 4.4 %) and 2.85 % (0.1 % to 4.5 %). Whereas the mean sucrose of honey sampled
from modern, transitional and traditional hives were 2.18 % (0 % to 4.5 %), 1.93 % (0.1 % to 4.4
%), and 1.03 % (0.1 % to 3.1 5), respectively. There was no significant difference observed both
due to agro-ecology and hive types of the present study.

Color: The overall average value of color of the samples used in the study area was 103.75 ± 2.89
mm which indicates amber color, with this the lowest color value obtained from the honey
sample acquired from the beekeepers with 75 mm which revealed light amber color; while the
highest color value was determined from retailers honey sample with 125 mm with dark amber
color. Higher finding reported by Tesfaye et al. (2016) greatest color of honey sample studied on
Pfund value was observed at Dinsho (140 mm Pfund) or dark amber and the lowest Pfund value
was observed at Adaba (20 mm pfund) or white color. The color of honey varies depending up on
the mineral content of honey, the darker the color the higher the mineral percentage of honey.

Quality parameters of honey of the study area had different correlation results between each
other. The moisture content of honey is positively and significantly correlated with acidity and
color content (P < 0.01). Moisture content has no correlation with the remaining parameters. The
HMF content is strong positive and significantly correlated electrical conductivity (P < 0.01) and
significant correlation (P < 0.05) with the free acidity. And also have weakly correlated with ash,
reducing sugar, sucrose and color and no correlation with pH. If honey is adulterated with
saturated sugar solutions, it will display greater conductance than pure honey as well for HMF
value. In fresh honeys there is practically no Hydroxy-methylfurfural (HMF), but it increases
upon storage, depending on the pH of honey and on the storage temperature (Bogdanov et al.
1999). Free acidity has significant correlation (p < 0.05) with the electrical conductivity. The
higher free acidity content, the higher the resulting conductivity (Bogdanov et al., 2002).Other
parameters had shown numerical strong and weak correlation between variables.

The questionnaire survey of this study was assessed the socio-demographic characteristics, KAP
of beekeepers on honey health benefits, major honey bee flora and challenges of beekeeping in
the study area. These characteristics related to the relative frequency distribution of household
heads by educational level, gender, age, beekeeping experience and livestock they owned.
Interestingly, the present study was further revealed that 15.20 % of the respondents have no
formal education which is in line with the reports of Ambaw and Teklehaimanot (2018), who
55
noted that 16 % of the interviewed beekeepers didn`t received either formal or informal
education. In the current finding the literacy level is 84.80%, which is higher than the national
average, i.e., 35.5% (Bogale et al., 2010), (Jenberie et. al, 2008) and (Behonegn, 2017) who have
reported that more than 60% and 62.5% of the sampled respondents of Amahara Region and
Sekota district, respectively were literate.Similarly, this finding is in agreement with Kinati et
al.(2011), who reported the majority of the interviewed beekeepers were educated in Gomma
district, south western Ethiopia.

The male to female ratio of the current study was 6.82: 1 (87.20 %) male. The aggressive
behavior of honeybee, time of harvest, burden of home job and overall house responsibility
limited the active participation of women‟s in beekeeping practices in the study area. Similarly,
Hartmann (2004), reported that traditionally beekeeping is mainly men‟s job in Ethiopia. The
traditional idea of the study area underscores beekeeping to be men`s job due to physical reasons.
This very limited number of female participation in beekeeping agrees with the findings of (Gela
and Negara, 2017) and (Abebe, 2011), who reported low level of women participation in
beekeeping. The very small number of female participation in beekeeping activities in the study
area is in line with the result of (Bihonegn, 2017) who reported 92.5% of total sampled
households (120) were male headed households while the rest were female headed in Tehulederie
district south wollo zone Amahara region, but incompatible with the report of (Fikru et.al, 2015)
who reported almost all of the respondents were males.

Average ages of the respondents were 43.31 ± 19.75 (with the range of 14 to 81 years). The
average age of the respondents of this study was nearly similar with the finding of Ambaw and
Teklehaimanot (2018), in Arsi zone, which is 42 years. Somewhat highest mean age result also
indicated by Getu and Birhan, (2014), reported mean age of respondents were 47.63 years in and
around Gondar. This proves that beekeeping is an important economic activity that can be
performed by all age groups, i.e. by younger and old people (Alemayehu, 2011).Differences in
beekeeping experience might be responsible to influence the attitude and adoption of new
beekeeping technologies (Wolters and Hussain, 2015). The beekeepers had an average
experience 7.66 (ranges from 3 to 26 years.) of beekeeping in the current study area. Higher
average experience and ranges of beekeepers experience was reported 12.47 years (ranges from 3
to 37 years) by Getu and Birhan (2014) in and around Gondar. The current finding of 96.80 % of
honeybee colony kept by traditional beehive is in line with the finding of (Tadesse and Kebede,
2014). According to Banchiamlak (2019), only about 10% of the honey produced in the Ethiopia
is consumed by the beekeeping households. The remaining 90% is sold for income generation

56
and of this amount, it is estimated that 80% is used for tej brewing. The same thing in this study
the respondents main purpose of keeping honey bees were for both household consumption and
income generation (88%), only for income generation (4%), and only for household consumption
(8 %) according to their need in this study. The main areas of Indigenous beekeeping knowledge
are hive construction from locally available materials, swarm catching, hive fumigation
materials, honey and swarming season identification, different medicinal values of honey,
identification of important honeybee floras, the medicinal value of honey and identification of
adulterated honey (Alemayehu, 2011). Beekeepers experiences in purpose of using honey
activities the deep indigenous knowledge of beekeepers which contributes to the development of
modern medicine varied from area to area and from agro-ecology to agro-ecology in the area.

Honey used as medicine has been limited due to lack of scientific report. In recent days, however,
there is resurgence. Its greatest medicinal potential is its application as topical agent to wounds
and skin infections. Honey has anti-inflammatory, immune boosting property, and exhibits broad
spectrum antibacterial activities (Motuma and Bekesho, 2016).Similarly, beekeepers of the
present finding used honey for the treatment of many diseases, symptoms and values but, some
of the respondents consume honey as foods without knowing the benefit of honey on the health
perspective. Infant botulism occurs in infants less than 1 year of age following ingestion of
spores in honey and syrup.The spores germinate in the gastrointestinal tract with toxin production
(Jackson, 2017). Of interviewed beekeepers 32.80 % aware on the honey side effects on infants
less than one year old while, 84 (67.20 %) hadn`t knew or heard before about the risk associated
with consumption of honey for infants less than one year old. In contrast to this, children, young
and old can alike take honey, without worrying any side effects (Kumar et al, 2010).

Dureja et al, (2003), stated that honey has been used as medicine in many cultures for a long
period of time. However, it has limited use in medicine due to lack of scientific report. In recent
days, honey is becoming acceptable as a reputable and effective therapeutic agent. Its beneficial
role has been endorsed to its antimicrobial, anti-inflammatory and anti-oxidant activities as well
as boosting of the immune system. In the present survey the knowledge of beekeepers honey
health benefits where mostly adopted 72 (64.29 %) from families, 34 (30.36 %) from neighbors
and 6 (5.36) were from other sources (book, radio, television and story). Knowledge of honey
health benefits have significant correlation with the beekeepers having long experience of
keeping honey colonies and age in the sampled study area. From the respondents interview the
aged group has more knowledge of honey health benefits and the highland beekeepers have 100
% knowledge of health benefits while 93.65 % from midland and 81.63 % from lowland agro-

57
ecologies were knew/ heard health benefits of honey from the total 125 beekeepers interviewed.
In other hand, this research revealed that educational background; of the respondents have almost
no relation with the knowledge, attitudes and practices of the honey health benefits.

In Ethiopia, people used traditional methods to treat both human and livestock diseases for
generations (Sori et al., 2004). Similarly, in the study area beekeepers used honey alone or by
mixing with other ingredients to treat many diseases, symptoms, disorders and values of peoples
and their livestock. Similarly, the main diseases, symptoms, disorders and values they treat
obtained from questionnaires were gastrointestinal tract, respiratory, skin, reproductive and they
used as cosmotics specially females. According to Dugassa et al,(2012) oral administration of
infusions, bolus, and other preparations used as route of administration to treat livestock and
humans by ethno-medicines. Likewise, in this finding the formulation of the honey for medicinal
value is bolus, syrup, juice, powder and infusion forms. The frequency and dosage of drugs given
is mostly as needed, from 1, 2 and 3 days as administered by the beekeepers local healers. The
species treated by the honey and additives were human being and livestock.

According to Mesfin et al,(2009), in Ethiopia 6500 melliferrous plant species of which more
1500 identified as bee forage. There are 58 National Forest Priority Areas in the Country that are
suitable for beekeeping .Of the total land mass of the country about 71% is suitable for fruit and
other crops growth which serve as source forage for the bees. Similarly, honeybee flora and trees
of the current study area are considered to be an essential indicator for potentialities for
beekeeping activity. A good beekeeping area is one in which honey and pollen plants grow
abundantly and with a relatively long blooming season. It requires the presence of appropriate
crops and plants to favor foraging. Many agro-forestry tree species were found in gardens of
farmers and were good sources of forage for the beekeepers (Gebru et al., 2015).As the results of
this survey indicated, the honey bee plants of the study area were trees, shrubs, herbs and
cultivated crops which are a source of nectar and pollen. Experienced beekeepers familiar with
the plants that produce nectar or/and pollen, when it blooms and how long they remain in
blooming.

Most beekeepers of the district knew the plant flora type and medicinal value of honey depending
on the color and test of honey. As indicated by Ambaw and Teklehaimanot (2018) in selected
districts of east and west arsi zone of Oromia region, different bee forage species were identified
by the respondents in local name as trees, shrubs and herbs. Accordingly the beekeepers of the
study area ststed that production of honey mostly comes from trees like Makanisa (Croton

58
macrostachys), Bargamo (Eucalyptus spp), Waddessa (Cordia Africana) and Baddessa
(MavlaVerticillate) usually flowers from march-July. Many beekeepers in the study area
explained that annual shrubs, crops and herbs like Hadaa (Bidens spp), Siddisa (Trifolium Spp)
and Nugi had yellow color and thick viscous honey. Similar report is presented in selected
districts of Arsi zone by Ambaw and Teklehaimanot (2018). Respondents of the stuy area stated
that the flowering season of this herbs, crops and shrubs were totally from September to
November, and the medicinal value of this honey was very higher than autumn harvested honey
from April to May.In this finding most important bee flowering shrubs, crops and weeds
flowered between September and November and most important plants flowered in March to
May and some in January the same finding is reported by Kumsa and Takele (2014) in Jimma
Zone. In Amuru district, the major honey harvesting season is between October and December.

The major constraints of beekeeping is the environmental condition which includes: honeybees‟
enemies, bee poisoning due to agrochemicals, lack of knowledge to manage bees and bee
products, lack of bee colonies and bees poisoning from plants (Tesfaye et al., 2017). In the same
manner beekeepers of the study district have faced with a number of challenges and difficulties
that affect their desired production. The challenges a rises from environmental factors that are
beyond the control of the beekeepers, while others have to do with poor marketing infrastructure
and management system. Beekeepers were interviewed to list the constraints in order of their
importance. The beekeepers of the study area experienced constraints of beekeeping 38.40 %
pests and predators, 20.80 % agrochemicals, 13.60 % Equipment and price of hive, 12 % disease,
8.80 % deforestation, 4 % market problem and 3.20 % lack of beekeeping knowledge.
Approximately similar report is presented by Bihonegn (2017) that, 60.74%, 46.67%, 45.93%,
27.41% and 22.22% of the respondents agree that the main reasons for the colony decline are
chemical application, lack of management, predators, pests and drought respectively.

59
6. CONCLUSION AND RECOMMENDATIONS

This study was conducted to evaluate honey quality, knowledge, attitude and practices of
beekeepers in Amuru district, Horo Guduru Wollega zone, Western Ethiopia. The main evaluated
parameters of honey samples were moisture content, HMF, pH, free acidity, ash, electrical
conductivity, reducing sugar, sucrose and color of honey analyzed and their evaluated mean
values were; 20.43 ± 1.32 %, 16.39 ± 2.68 mg/kg, 3.92 ± 0.07, 36.67 ± 2.13 meq/kg, 0.25 ± 0.14
%, 0.68 ± 0.3 mS/cm, 73.08 ± 0.92 %, 1.80 ± 0.35 % and 103.75 ± 2.89 mm respectively. The
value of moisture content, HMF, pH, free acidity and color significantly (p < 0.05) influenced by
sources of the sample (i.e. beekeepers and retailers). Whereas, the value of moisture content,
HMF and free acidityis significantly (p < 0.05) affected among agro-ecologies. Statistically there
was no significance difference (p > 0.05) among hive types in terms of all analyzed
parameters.The moisture content value of honey sampled declared significant positively
correlated with free acidity and color r = 0 .63 and r = 0.53 respectively, (p < 0.01).While, HMF
value has significant (p < 0.01) positive correlation with Electrical conductivity value r = 0.77
and significantly (p < 0.05) correlated with free acidity r = 0.45. Free acidity has significant (p <
0.05) correlation with electrical conductivity value r = 0.41.

The beekeepers have developed indigenous KAP of keeping bees passing from generation to
generation in the study area. These indigenous beekeeping knowledge are hive construction from
locally available materials, honey health benefits, major constraints of beekeeping, swarming
season identification, identification of important honeybee floras and flowering season of bee
flora.Most of the respondents were keeping honey colony for the purpose of both consumption
and income. Majority of the respondents (89.60% knew or heard the health benefits of honey and
practiced to treat many of the human and animal disease and symptoms. The major constraints
identified for beekeeping in the study area are pests and predators, indiscriminate agro-
chemicals disease, lack and expensiveness of equipment, deforestation and other factors. In
conclusion results revealed that honey produced in Amuru district is of good quality and can
meet the national and international market demands. This result provides baseline data on pattern
of honey quality and production and identifies perception gaps that can be used as reference for
future in the study area.

60
Based on the research work conducted in the study area, the following recommendations were
forwarded:

 To improve the honey quality defects associated with poor management, harvesting and
handling in the study district specially by honey retailers, there is a need to provide a
practical training to local beekeepers and retailers about proper ways of handling,
processing, packaging and sale of honey, moreover, facilitating supply of quality
apicultural equipment is crucial.
 Address the skill gap on bee colony management and post harvest handling of hive
products, further consistent practical training on bee and bee products management for
community is recommended.
 Now a days, there is alarming time to search 3rd generation medicine due to newly
emerging and re-emerging disease, so government and non-governmental sectors should
have to promote KAP of beekeepers and other social practices.
 Female beekeepers should be empowered to participate effectively in beekeeping
activities by offering practical training, organizing in different cooperatives and providing
credit service by governmental and non-governmental institutions.
 The government should integrate multidisciplinary experts with the traditional healers to
develop modern pharmaceutical products which may solve world‟s current straining. e.g.
the indigenous knowledge on honey health benefits

 Farmers of the study district were using numerous chemicals for the protection of crop
during day time when honeybees foraging, the toxicity effect of each chemical on
honeybee is too high, so, the farmer should spray the chemical during night time.

 Further study should be recommended to assess honey production and physicochemical

properties of honey in the study area.

61
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8. ANNEXES

Annex 1. Sample collecting sheet

Source

Agro-ecology
Amount (kg )

and Lowland
Sample code

Type of bee

(High, Mid

Remarks
Beekeep

Retailer
Type of
Sample

District
(HS)

hive
No.

ers
1 Honey 0.5 kg
2 >> >>
3 >> >>
4 >> >>
5 >> >>
6 >> >>
7 >> >>
8 >> >>
9 >> >>
10 >> >>

Annex 2. Questionnaire recording sheet

No Name Zone District Kebele Age Sex Educat Agro Type of hive
. of ional ecology
respond backgr
ents ound

Mod
Trns
Trd
Ml
Hl

Ll

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15

71
Annex 3. Procedure for honey moisture determination
1. Preparation of the sample
2. Determine the refractive index of the test sample using a refractometer at a constant
temperature near 20 0C.
3. Convert the reading to moisture content (percent by mass) using the table.
4. If the determination is made at a temperature other than 20 0C, convert the reading to
0
standard temperature of 20 C, according to the temperature corrections quoted.

Figure 1: Abbe refractometer

72
Annex 4. Conversion factor for the estimation of moisture content from refractive index data

Water RI (20°C) Water RI (20°C) Water RI (20°C) Water RI (20°C)

content content % content content
% % %
13.0 1.5044 16.0 1.4966 19.0 1.4890 22.0 1.4815

13.2 1.5038 16.2 1.4961 19.2 1.4885 22.2 1.4810

13.4 1.5033 16.4 1.4956 19.4 1.4880 22.4 1.4805

13.6 1.5028 16.6 1.4951 19.6 1.4875 22.6 1.4800

13.8 1.5023 16.8 1.4946 19.8 1.4870 22.8 1.4795

14.0 1.5018 17.0 1.4940 20.0 1.4865 23.0 1.4790

14.2 1.5012 17.2 1.4935 20.2 1.4860 23.2 1.4785

14.4 1.5007 17.4 1.4930 20.4 1.4855 23.4 1.4780

14.6 1.5002 17.6 1.4925 20.6 1.4850 23.6 1.4775

14.8 1.4997 17.8 1.4920 20.8 1.4845 23.8 1.4770

15.0 1.4992 18.0 1.4915 21.0 1.4840 24.0 1.4765

15.2 1.4987 18.2 1.4910 21.2 1.4835 24.2 1.4760

15.4 1.4982 18.4 1.4905 21.4 1.4830 24.4 1.4755

15.6 1.4976 18.6 1.4900 21.6 1.4825 24.6 1.4750

15.8 1.4971 18.8 1.4895 21.8 1.4820 24.8 1.4745

25.0 1.4740

Source: (Wedmore 1955)

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Annex 5. Procedures for hydroxyl-methyl furfural
1. Accurately weigh 5 g honey in small beaker and transfer with total of 25 ml distilled water to
50 ml volumetric flask
2. Add 0.50 Carrez solution I, mix, add 0.50 ml Carrez solution II, mix, and dilute to volume
with distilled water drop of alcohol may be added to suppress foam.
3. Filter through filter paper, discarding the first 10 ml filtrate.
4. Pipette 5 ml filtrate into each of two 18 X 150 mm test tubes.
5. Add 5.0 ml distilled water to one tube (sample) and 5.0 ml NaHS03 solution to other.
6. Mix well by using Vortex mixer and determine.
7. Absorbance of sample against reference at 284 and 336 nm in 1 cm cells.
8. If absorbance is greater than 0.6, dilute sample solution with water and reference solution with
0.1% NaHS03 solution to same extent and correct absorbance for dilution.

Annex 6. Dilution of sample and reference solutions carried for estimation of HMF

Addition to test-tubes Sample solution (in ml) Reference solution (in ml)

Initial solution 5.0 5.0

Water solution 5.0 -

Sodium bisulphate (0.2 %) - 5.0

Source: IHC, 2009

Figure 2. Sample solution and reference solution

74
Dilution (D) =

Calculation and expression of result:

HMF expressed as mg/kg = (A284-A336) x 149.7 x 5 x D/W.
A284= Absorbance at 284 nm.
A336 = Absorbance at 336 nm.
149.7 = 126 x 1000 x 1000/16830 x 10 x 5.
126 = Molecular weight of HMF.
16830 = Molar absorptive and HMF at 284 nm.
10 = Conversion of g into mg.
1000 = Conversion of g into kg.
5 = Theoretical nominal sample weight.
D = Dilution factor (in case dilution is required)
W= Weight in g of honey sample

Annex 7. Procedure for Acidity and PH

1. Preparation of test sample; weigh 10 gram of honey accurately and dissolve it in 75ml in
distilled water
2. The test sample is titrated against carbonate free 0.1N sodium hydroxide solution using 4-5
drops of phenolphalein indicator
3. The end point colour should persist for 10 seconds
4. For dark cooled sample a smaller weight shall be taken
5. As an alternative a PH meter may be used and the sample titrated to PH 8.3
6. Expression of the result

Acidity=10V
Where V = the volume of 0.1M NaOH used and 10 is the amount of honey sample

75
Figure 3. pH meter and Magnetic strirer

Annex 8. Procedures for determining ash content

1. Weigh 5 to 10 grams Honey accurately into ignited and pre-weighed platinum, silica or
porcelain crucible (shiinii).
2. Gently heat in muffle furnace until the sample is black and dry.
3. An infra-red lamp or Bunsen burner can also be used to char the sample before inserting in to
the muffle furnace.
4. If necessary; a few drop of olive oil may be added to prevent frothing.
5. The sample is then ignited at 600oc to constant weight.
6. The sample is cooled in desiccator before weighing.

7. Finally weigh and calculate the percentage.

 Percent ash in g/100g honey was calculated using the following formula, following the
procedure of (Bogdov, 2009 )

Where; mo = weight of honey take m1 = weight of dish + ash, m2 = weight of dish. Proposed
ash content not more than 0.6 % for normal honey.

76
Figure 4. Hot plate and muffle furnace

Annex 9: Procedure of determining reducing sugar and sucrose by HPLC Method

A. Reagent
 Chemicals of analytical purity grade were used.
1. Distilled water HPLC (purchased from the Finfine Kirkos Heparin Trading private
limiting Company) 4500 ETB (2.5 Litre).
2. Methanol for HPLC (from lab.)
3. Acetonitrile for HPLC. Purchased from Finfine, Kirkos, Heparin Trading private limiting
Company 5200 ETB (2.5 Litre). Warning: Acetonitrile is a dangerous substance
laboratory safety guidelines on dangerous substances at work were used
4. Eluent solution for the HPLC. Mix 80 volumes of acetonitrile with 20 volumes of water
and degassed prior to use.
5. The standard substances: fructose, glucose and sucrose obtained from lab.
 fructose: 2.000 g
 glucose: 1.500 g
 sucrose: 0.250 g
B. Equipments
Sample vials, Ultrasonic bath, Calibrated flasks, volume 100 ml, 25-ml-pipette, Membrane filter
for aqueous solutions, pore size 0.45 μm, Filter holder for membrane filters with suitable syringe,
High Performance Liquid Chromatograph consisting of pump, sample applicator, temperature
regulated RI-detector thermostated at 30o C*, temperature regulated column oven at 300C,
integrator, analytical stainless-steel column, silica gel with 5-7 μm particle size. Before use, a
system suitability test to ensure all the sugars can be separated was carried out.

77
* Note: the chromatography can be carried out at room temperature without influence on the
results of the sugars, determined by the present method.
C. Procedures
Preparation of the sample solution
1. Weigh 5g of honey into a beaker and dissolve in 40 ml water.
2. Pipette 25ml of methanol into a 100ml volumetric flask and transfer the honey solution
quantitatively to the flask.
3. Fill to the mark with water.
4. Dissolve the amounts detailed of the standards in approximately 40ml water and transfer
quantitatively to the flask and fill to the mark with water
5. Pour through a membrane filter and collect in sample vials.
6. Store as for the standard solution.
 High Performance Liquid Chromatography (HPLC)
-After a column of the type described above is used, the following conditions have been found
to give satisfactory separation.
 Flow rate: 1.3 ml/min
 mobile phase: Acetonitrile:water (80:20, v/v)
 column and detector temperature : 300C
 sample volume: 10 μl
Note: Identical volumes of sample and standard solution should be injected.
D. Calculation and Result Expression
The honey sugars are identified and quantified by comparison of the retention times and the peak
area of the honey sugars with those of the standard sugars. The mass percentage of the sugars,
W, to be determined of fructose, glucose, etc..and maltose in g/100g is calculated according to
the following formula (external standard procedure):
W = A1 x V1 x m1 x 100/ A2 x V2 x m0
Where,
A1 = Peak areas or peak heights of the given sugar compound in the sample solution, expressed
as units of area, length or integration.
A2 = Peak heights of the given sugar compound in the standard solution, expressed as units of
area, length or integration.
78
V1 = Total volume of the sample solution in ml
V2 = Total volume of the standard solution in ml
m1 = Mass amount of the sugar in grams in the total volume of the standard (V2)
m0 = sample weight in g. The final result is rounded to one decimal place.

Figure 5. Honey sample processing in HPLC machine to calculate reducing sugar and sucrose

Annex 10. Procedures for color determination

1. Homogeneous honey samples free of air bubbles was transfer into a cuvette (transparent
tubularLaboratory vessel) with a 10-mm light path until the cuvette is approximately half full.
2. The cuvette was inserted into a color photometer Pfund honey color grader (No. 0061, made in
USA)
3. The color grades were expres sed in millimeter (mm) Pfund grades compared to an analytical
gradeglycerol standard following the procedure of IHC (2009).
Annex 11. USDA color standard

Standard P-fund Color Grader (mm)

Water white 0-8
Extra white ≥ 8-17
White ≥17-34
Extra light amber ≥ 34-50
Light amber ≥ 50-85
Amber ≥ 85-114
Dark amber ≥ 114
Source: (IHC, 2009)

79
Figure 6. Pfund color grader

80
Annex 12. The results of all the honey quality parameters analyzed in the study area (N = 24)

Source of honey Quality Parameters

samples
Agro- Hive types MC HMF pH FA Ash EC Rs AP Color
ecology (%) (mg/kg) (meq/cm) (%) (mS/cm) (%) (%) (mm)

Highland Modern 19.4 0 4.1 22 0.2 0.4 73 1 101

Transitiona 1.4 3.9 33 0.1 0.3 74 1.4 108
l 20.4
Traditional 21.8 2.1 4.1 30 0.1 0.3 69 0.1 102
Midland Modern 20.2 3.2 4 33 0.3 0.6 78 0.1 96
Modern 20.4 1.8 4 34 0.2 0.5 74 3.4 108
Modern 21.0 2.8 4.1 32 0.1 0.3 71 0 109
Transitiona 6.6 3.9 40 0 0.2 78 0.1 109
l 20.4
Transitiona 6 3.9 43 0.2 0.4 71 4.4 75
l 19.6
Transitiona 7.3 3.9 42 0.5 1 71 0.4 108
l 20.0
Traditional 20.6 9.8 3.7 48 0.4 0.9 70 2.7 78
Traditional 21.0 8.8 4 31 0.5 0.3 71 0.1 107
Traditional 21.2 11 3.9 43 0.4 0.7 80 0.1 108
Lowland Modern 19.0 16.4 4.2 14 0.3 0.7 69 4.5 102
Modern 19.2 12.3 4.8 24 0.2 0.7 66 4.1 110
Transitiona 26.7 4.3 26 0.3 0.8 76 2.9 105
l 18.0
Transitiona 23.1 4 29 0.1 0.8 80 2.4 79
l 19.0
Traditional 19.2 29.1 3.8 46 0.3 0.8 68 3.1 78
Traditional 18.2 25.3 4.2 24 0.5 0.9 79 0.1 98
Honey Retailer 22.4 39.2 3.1 49 0.4 0.9 79 0.1 125
retailers Retailer 21.8 38.6 3 55 0.2 0.9 78 0 129
Retailer 21.0 36 3.8 45 0.2 1 75 2.5 118
Retailer 21.0 30.5 3.6 47 0.1 1 67 3.5 112
Retailer 22.6 24.1 3.8 46 0.3 1 68 4.8 107
Retailer 22.8 31.3 3.9 44 0.1 1 69 1.5 118

81
Annex 12. Results of Univariable GLM analysis of honey samples parameters

Independent variable Dependent variable No. of tested p-value

Agro-ecology MC (%) 18 .000
HMF (mg/Kg) 18 .000
pH 18 .073
FA (meq/Kg) 18 .031
Ash content (%) 18 .306
EC (mS/cm) 18 .295
RS (%) 18 .838
As (%) 18 .124
Color (mm) 18 .642
Source MC (%) 24 .000
HMF (mg/Kg) 24 .000
pH 24 .001
FA (meq/Kg) 24 .001
Ash content (%) 24 .526
EC (mS/cm) 24 .528
RS (%) 24 .800
As (%) 24 .675
Color (mm) 24 .002
Type of hive MC (%) 18 .449
HMF (mg/Kg) 18 .318
pH 18 .148
FA (meq/Kg) 18 .097
Ash content (%) 18 .114
EC (mS/cm) 18 .156
RS (%) 18 .466
As (%) 18 .496
Color (mm) 18 .437

82
Annex 13. Quality of honey sample of the study area as compared to QSAE, EU and CAC:

Parame N The current result of the National and International institutions and their
ters study area standards
Mean ± SD Max. Min. QSAE EU CAC World FAO/WHO
Moistur 24 20.4± 1.32 22.8 18 17.5-21 < 21 < 21 18-23 21-23
e%
HMF 24 40 < 40 < 60 40-80 ≤ 80
(mg/kg 16.39 ± 2.68 39.2 0
)
PH 24 3.92 ± .07 4.8 3 - - 3.2-4.5 -
FA(me 24 36.67 ± 2.13 55 14 40 < 40 < 50 5-54 40
q/kg)
Ash 24 0.25 ± 0.14 .50 0 0.6 <0.6 < 0.6 0.25 – 1 0.6 – 1
(%)
EC 24 0.6 ± 0.3 1 0.1 <0.6 <0.8 <0.8 .22-1.52 -
mS/cm
RS (%) 24 73.08± 0.92 80 66 65 < 65 < 65 60 – 70 ≥ 65
Sucrose 24 1.80 ± 0.35 0 4.8 10 <5 <5 3-10 5-10
(%)
Color 24 103.75±2.89 129 75 - - - - -
(mm)
Source : CAC (2001); QSAE (2009); EU Council (2002) and QSAE (2000)

QSAE, quality and Standards Authority of Ethiopia; CAC ; CAC; EU; European Union; meq;
milli equivalent

83
Annex 14. Questionnaires used in the study
 Jimma University College of Agriculture And School of Veterinary Medicine
 Department of Veterinary Public Health
Evaluation of Honey Quality and the KAP of beekeepers In Amuru district Of Horo
Guduru Wollega Zone, Oromia, West Ethiopia.
o This questionnaire is prepared for an academic purpose for the fulfillment of MSc degree
in Veterinary Public Health.
o Dear respondent: This questionnaire is designed to collect necessary information to
assess beekeepers practices on health benefits of honey and the beekeeping system in
Amuru district; Western Ethiopia.
o So your response to the question has great contributions. Therefore you are politely
requested to give the accurate and necessary information.

I. Direction for Enumerator

 Please greet them and introduce yourself patiently to the beekeepers before starting the
interview.
 Tell the respondents politely about the aim and purpose of the study
 Try to start with easy questions
 Use pencil to tick on Yes or No and to write broad idea on provided space
 Create sufficient awareness for beekeepers regarding with questions to get accurate data
 Write down properly the idea provided by the beekeepers
 Ask the questions by using the language that beekeepers can understand and know easily
A. Interview guideline for district Office of Livestock Resource Development and Fishery

1. What is the agro-ecology of the district?

A. High land B. Mid land C. Low land D. All E. Other____________________________
2. How many kebeles are high, mid and low land agro-ecology?
Highland__________ Midland__________Lowland________________________________
3. Number of beekeepers in the district in 2011 E.C. ___________________________________
4. Total number of beehives in the district in 2011 E.C._________________________________

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 A.Traditional______________ B.Transitional____________ c. Modern__________________
5. What is the average productivity of the different hives (Kg/hive/harvest) in the the district?
Traditional ____________, Transitional__________Modern___________________________
6. Which kebeles have high beekeeping practices? ____________________________________
Why?_________________________________________________________________
8. At what season honey mostly harvested in the district? _______________________________
9. What is the general perspective of the beekeeping activities in the district?
__________________________________________________________________
10. How is the potential and what is the good opportunities for beekeeping in the district
?__________________________

B. Questionnaires for Beekeepers

1. Socio-demographic history of beekeepers

 Name of beekeeper______________________________Zone________District________
Kebele________Village_______________Agro-ecology__________
 Sex: a. Male b. Female
 Age: a. 14-25 b. ≥25-65 c. ≥ 65
 Educational background A. Illiterate B. Elementary school C. High school D.
College/University

2. Hive type and honey production potential

2.1. What type of bee hive do you have?

Hive type  or x Amount
Modern
Transitional
Traditional

2.2.When did you start beekeeping? A. < 5 year B. ≥ 5-10 years C. > 10 year
2.3. At what time do you harvest honey? A. day B. night C. specify if any other____________
2.4. If your answer in question number 2.3. Is A or B why at that time? ___________________

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2.5. The amount of honey produced

No. Type of hive Frequency of Average of honey Total amount of

harvesting/year harvested honey produced
(season/hive/ kg) hive/year in Kg
1 Modern
2 Transitional
3 Traditional

2.6. What are peak honey producing months_________________________________________?

3. Honey health benefits KAP

3.1. For what purpose you keep honey bee colony?

A. both for income and consumption B. only for household consumption C. only for income
3.2. Do you know or heard the health benefits of honey? A. yes B. no
3.3. If your answer on question no.3.2. Is yes;
A. For what disease, disorders, symptoms / values you use? __________________________
B. Formulation: ____________________________________________________________
C. Route of administration: ___________________________________________________
D. Frequency; ______________________________________________________________
E. Dosage: ________________________________________________________________
F. Species it treats; _________________________________________________________
3.4. From where do you get the knowledge and practices of honey health benefits?
A. From family B. from neighbors C. from School D. Others ______________
3.5. Have you heard before the honey side effect on child < 1 year old? A. yes B. no
3.5. If your answer on question no. 3.4. Is yes, what is the side effect?
______________________________________________________________________________
4. Honey Bee Plants and Flora
4.1. What are the major honey bee plants or floras in your area and their flowering season?

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 No. Local name of honey bee plants/floras Flowering season
1
2
3
4
5
4.2. What are the peak flowering seasons of the year? _________________________________
5. Factors Influencing Beekeeping system in the study area
5.1. What are the factors that influence beekeeping in your area?
A. Predators and Pests B. Agro-chemicals C. Deforestation D. Lack of beekeeping knowledge
E. Lack Market F. lack of equipment and prices of hive
5.2.Is there a trend of using agro-chemicals in your locality for crop production? A. yes B. no
 Name of Enumerator:_____________________________
 Signature: ______________________________________
 Date of interview:________________________________

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