FINAL YEAR RESEARCH PROPOSAL

ASSESSING THE COMPARATIVE EFFICACY OF SELECTED MEDICINAL PLANTS

AGAINST TICK LARVAE IN NAMALU SUB-COUNTY, NAKAPIRIPIRIT DISTRICT

BY
LEESE ADAM
BU/UP/2021/0163
SUPERVISOR: PROF. OLILA DEO

MAY 2024

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DECLARATION
I leese Adam, declare that this information is original and has never been submitted to any higher
institution of learnig and has been developed for the award of degree in animal production and
management of Busitema university.
SIGNATURE……………………….
DATE……………………………..

APPROVAL
This research proposal is submitted with approval by my research supervisor:
Prof. Olila Deo
Sign……………………………

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Date …………………………..

DEDICATION
I dedicate this work to my parents Mr. Benga Titus and Mrs. Nait lucy and all my brothers and
sisters for their support through my university education. My sincere thanks to my supervisor
Prof. Olila Deo for his guidance towards the development of my proposal.

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ACKNOWLEDGMENT
First and foremost, I would like to express my deepest gratitude to God for granting me the
strength, perseverance, and wisdom to complete this research project.

I would also like to extend my heartfelt appreciation to my parents, Mr. Benga Titus and Mrs.
Nait Lucy and all my siblings for their unwavering love, support, and encouragement throughout
my academic journey have been instrumental in my success.

My sincere thanks go to my esteemed research supervisor, Prof Olila Deo, for their invaluable
guidance and mentorship. His expertise, patience, and constructive feedback have shaped this
research and pushed me to achieve my full potential.

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TABLE OF CONTENTS
DECLARATION...............................................................................................................................................i
APPROVAL....................................................................................................................................................ii
DEDICATION................................................................................................................................................iii
ACKNOWLEDGMENT...................................................................................................................................iv
CHAPTER ONE: INTRODUCTION...................................................................................................................1
1.0. BACKGROUND.......................................................................................................................................1
1.2 Problem Statement:...............................................................................................................................1
1.3.1 General Objective:..............................................................................................................................2
1.3.2 Specific objectives...............................................................................................................................2
1.4 Research questions................................................................................................................................2
1.5 Significance of the Research Topic:........................................................................................................2
1.6 Justification of the Research:.................................................................................................................3
1.7 Scope of the Research Topic:.................................................................................................................3
CHAPTER TWO: LITERATURE REVIEW..........................................................................................................4

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2.0 Literature review...................................................................................................................................4
2.1 Economic importance of ticks...........................................................................................................4
2.2 Prevalence Studies:............................................................................................................................5
2.2.1 Karamoja prevalence studies..........................................................................................................5
2.2.2 Dominant Tick Species:..................................................................................................................6
2.3 Tick Resistance: A Global Threat with Regional and National Nuances............................................6
2.3.1 Global Trends:................................................................................................................................6
2.3.2 Regional Trends:.............................................................................................................................7
2.3.3 National Trends:.............................................................................................................................7
2.3.4 Possible Causes of Resistance:.......................................................................................................7
2.3.5 Potential Consequences:.................................................................................................................7
2.4 Plants as sources of pesticides/acaricides..........................................................................................8
2.4.1 Ricinus communis..........................................................................................................................8
2.4.2 Ocimum basilicum..........................................................................................................................8
2.4.3 Carica papaya.................................................................................................................................8
2.4.4 Pyrethroids......................................................................................................................................8
2.5 Acaricidal assays...............................................................................................................................9
2.5.1 Contact toxicity assays:................................................................................................................10
2.5.2 Fumigation assays:.......................................................................................................................10
2.5.3 Repellency assays:........................................................................................................................10
2.5.4 Residual activity assays:...............................................................................................................10
2.6 Life cycle and behavior of ticks.......................................................................................................10
2.6.1 Life Cycle:....................................................................................................................................10
2.6.2 Feeding Habits:.............................................................................................................................11
2.6.3 Behavior:......................................................................................................................................11
2.7 In vitro acaricidal testing.................................................................................................................12
3.0 Methodology.......................................................................................................................................12
3.1. Study area...........................................................................................................................................12
3.2 Study design...............................................................................................................................13
3.3 Research Design:..................................................................................................................................13
3.4 Research Approach:.............................................................................................................................14

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3.5 Sample Size Selection:.........................................................................................................................14
3.6 Study Population:................................................................................................................................14
3.7 Identification of medicinal plants........................................................................................................14
3.8 Transportation and Storage of Selected Plant Samples:......................................................................15
3.8.1 Transportation:.................................................................................................................................15
3.8.2 Storage:.............................................................................................................................................15
3.9 Operational design...............................................................................................................................16
3.4. Data analysis........................................................................................................................18
3.10 Ethical Considerations:......................................................................................................................18
Questionnaire............................................................................................................................................20
REFERENCES..............................................................................................................................................23

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CHAPTER ONE: INTRODUCTION

1.0. BACKGROUND

Ticks serve as vectors for various diseases affecting both humans and animals, posing significant
challenges to livestock management and public health. These parasites not only directly harm the
health of livestock but also transmit diseases such as East Coast fever and tick-borne
encephalitis, threatening human well-being. Moreover, ticks induce economic losses by reducing
livestock productivity and necessitating costs. Different groups of chemicals have been used
however there is need to explore more natural remedies to ticks .(Srivastava et al., 2008)
Traditionally, the control of ticks has heavily relied on synthetic acaricides, despite concerns over
their adverse environmental effects and the development of resistance among tick populations.
To address these challenges, there is a growing interest in exploring alternative and sustainable
tick management strategies, with medicinal plants emerging as promising candidates due to their
pesticidal properties.(Mutavi et al., 2021)
Medicinal plants, renowned for their diverse biological activities, offer potential solutions for
environmentally friendly tick control. With their natural compounds exhibiting repellent and
acaricidal effects, these plants represent an opportunity to develop sustainable tick management
practices while minimizing environmental impact and safeguarding human and animal health.
(Ghosh et al., 2011)
The lack of comprehensive knowledge on the extraction methods and bioactive compounds of
medicinal plants adds to the uncertainty surrounding their utility in tick control. Understanding
the optimal extraction techniques to obtain potent acaricidal compounds and assessing their
safety for livestock, wildlife, and the environment are essential aspects that require further
investigation.
Therefore the study aims to explore indigenous knowledge on medicinal plants used against ticks
in Namalu sub county and to scientifically validate their efficacy against ticks.

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1.2 Problem Statement:
Despite the rich biodiversity and traditional knowledge surrounding medicinal plants in Namalu
Sub County, there exists a significant gap in understanding their potential role in tick
management. While local communities may have historically utilized certain plants for pest
control, there is a lack of scientific validation and systematic exploration of their efficacy against
ticks. This gap hampers the development of sustainable and culturally relevant tick control
strategies.(Liu et al., 2023)
The current reliance on synthetic acaricides exacerbates environmental degradation and health
risks, necessitating the exploration of alternative approaches. However, the specific medicinal
plants indigenous to Namalu Sub County with potential acaricidal properties remain largely
unidentified and unexplored. Without empirical evidence on the efficacy, safety, and application
methods of these plants, their integration into tick management practices remains speculative.
(Kemal et al., 2020)
Furthermore, the lack of comprehensive knowledge on the extraction methods and bioactive
compounds of medicinal plants adds to the uncertainty surrounding their utility in tick control.
Understanding the optimal extraction techniques to obtain potent acaricidal compounds and
assessing their safety for livestock, wildlife, and the environment are essential aspects that
require further investigation.
Additionally, there is a dearth of research on the long-term sustainability and ecological impact
of integrating medicinal plants into tick management strategies. The potential consequences of

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 widespread plant harvesting for pest control purposes on biodiversity, ecosystem stability, and
traditional knowledge systems remain largely unexplored.(Mkwanazi et al., 2021)
Addressing these knowledge gaps is crucial for developing evidence-based tick control
interventions that align with local practices and environmental sustainability goals. By bridging
the divide between traditional knowledge and scientific inquiry, this study aims to contribute to
the development of effective, culturally relevant, and environmentally sustainable tick
management strategies in Namalu Sub County.
.

1.3.1 General Objective:

 To assess the efficacy of selected medicinal plant extracts against hard ticks

1.3.2 Specific objectives

 To collect indigenous information on plants used against ticks in Namalu sub county
 To determine the comparative vitro efficacy of selected medicinal plants extracts against
ticks.

1.4 Research questions

I. Which medicinal plants are traditionally used by the community in Namalu sub-county
against ticks?
II. What is the comparative efficacy of selected medicinal plant extracts against ticks?

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1.5 Significance of the Research Topic:

Limited Empirical Data: Traditional use of medicinal plants for pest control in Namalu Sub
County may lack empirical validation. The research will provide scientific evidence to confirm
or challenge these traditional claims, enhancing credibility.

Comparative Efficacy: Few studies may have systematically compared the effectiveness of
multiple medicinal plants against tick larvae. The research will fill this gap by providing a
comparative analysis, identifying which plants are most effective and under what conditions.

1.6 Justification of the Research:

Scientific and Practical Validation: The research addresses gaps in scientific understanding by
validating traditional knowledge and identifying effective medicinal plants for tick control. This
will provide a solid foundation for integrating traditional practices with modern pest
management strategies.

Addressing Local Needs: By focusing on locally relevant plants and pest management
challenges, the study offers practical, cost-effective, and environmentally sustainable solutions
tailored to the specific needs of Namalu Sub County.

Economic and Educational Impact: The study promotes local economic development, supports
sustainable agricultural practices, and provides educational opportunities that empower local
communities and enhance their capacity to manage tick infestations effectively.

1.7 Scope of the Research Topic:

Geographic Scope: The primary focus of the research is Namalu sub-county in Nakapiripirit
District in the northeastern region of Uganda. The study will concentrate on the local flora and
traditional knowledge relevant to this specific geographic area.
Medicinal Plant Selection and Identification: The research will involve a systematic survey of
indigenous medicinal plants in Namalu sub-county known for their pesticidal properties or
traditionally used for pest control purposes. This survey will include consultations with livestock
farmers to identify potential candidate plants. The selection process will prioritize species with
documented traditional use and anecdotal evidence of efficacy against ticks.
Acaricidal Testing: Laboratory experiments will be conducted to assess the acaricidal efficacy of
extracts from selected medicinal plants against ticks. The study will employ established in vitro
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assays to evaluate the mortality rate and repellent effects of plant extracts on tick populations.
These experiments will be conducted at a designated research facility within Namalu sub-county,
equipped with the necessary laboratory equipment and personnel.
Timeline: The research will be conducted over a specified timeframe, with distinct phases for
plant selection, extraction, and acaricidal testing. The duration of each phase will be determined
based on logistical considerations, resource availability, and the complexity of the research
protocols. A timeline of approximately 2-3 months is anticipated for completing the entire
research project, including data analysis and dissemination of findings.

CHAPTER TWO: LITERATURE REVIEW

2.0 Literature review

2.1 Economic importance of ticks

Ticks pose a significant economic threat in Uganda, with annual losses exceeding USD 1.1
billion(Kasaija et al., 2021) primarily attributed to the transmission of economically important
pathogens. East Coast fever (ECF), caused by Theileria parva and transmitted by Rhipicephalus
appendiculatus, stands out as a major contributor to these losses. Beyond direct cattle mortality,
ECF leads to reduced productivity, treatment expenses, and long-term socioeconomic impacts for
farmers. Additionally, ticks transmit various other diseases, further exacerbating economic
burdens through treatment costs and trade restrictions. The labor and resources invested in tick
control measures add to the economic strain. Addressing tick infestations and tick-borne diseases
requires holistic strategies encompassing research, interventions, and policy measures to
safeguard Uganda's agricultural sector and ensure food security for its population(Kasaija et al.,
2021)
80% of the world's cattle population is affected by ticks, beyond being vectors for diseases that
impact livestock health ticks also pose economic burdens on farmers worldwide. Alternative
control methods while utilized often face resistance necessitating exploration of other
alternatives. The occurrence of ticks across diverse climates and regions means that livestock in
many parts of the world are at risk, putting a need for concerted efforts in research, collaboration
and holistic approaches to tick management(Hurtado and Giraldo-Rios 2019)
The statement from the Uganda Presidential Advisory Committee in 2017 highlights the
significant economic impact of tick infestations and tick-borne diseases on the animal resource
industry, despite its substantial contribution of 17% to the country's gross domestic product
(GDP). In Uganda, as in many other regions with rural economies, livestock farming plays a
crucial role in providing livelihoods for rural communities and contributing to national economic
development. However, the prevalence of ticks poses a persistent challenge to this sector, leading
to reduced productivity, increased mortality rates, and substantial economic losses for farmers
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and the wider economy. These losses are incurred through decreased livestock productivity,
including reduced meat and milk production, as well as costs associated with disease treatment,
tick control measures, and potential trade restrictions due to disease concerns. As such,
addressing tick infestations and tick-borne diseases is essential not only for safeguarding animal
health and welfare but also for sustaining the economic viability of the animal resource industry
and supporting broader socio-economic development goals in Uganda. (Uganda Presidential
Advisory Committee 2017)
In Uganda ticks and tick-borne diseases cause a 75% loss, attributed to ticks and tick-borne
diseases in Uganda underscores the profound impact of these parasites on the country's
agricultural sector and economy. This level of loss reflects a multifaceted challenge
encompassing reduced livestock productivity, increased mortality rates, and substantial economic
burdens on farmers and the wider economy. Livestock farming is a cornerstone of Uganda's rural
economy, providing livelihoods for a significant portion of the population and contributing to
national food security and economic development. However, the prevalence of ticks and the
diseases they transmit pose persistent obstacles to the sector's growth and sustainability. The
economic losses incurred include decreased meat and milk production, costs associated with
disease treatment and tick control measures, and potential trade restrictions due to disease
concerns. Addressing this issue requires coordinated efforts involving research, extension
services, veterinary care, and policy interventions aimed at improving livestock health,
enhancing tick control strategies, and mitigating the economic impact of tick-borne diseases on
Uganda's agricultural sector(Ocaido et al., 2009)

2.2 Prevalence Studies:

2.2.1 Karamoja prevalence studies

Several studies have assessed tick prevalence in Nakapiripirit District and surrounding areas:
A study reported a 72.2% prevalence of `R. e. evertsi` on cattle in Karamoja, highlighting its
widespread distribution and role in ECF transmission, indicating a widespread distribution of this
tick species. This finding underscores the significant role of R. e. evertsi in the transmission of
East Coast fever (ECF), a severe and often fatal disease affecting cattle, caused by the protozoan
parasite Theileria parva. Understanding the prevalence and distribution of such tick vectors is
pivotal for devising effective control measures against ECF. Strategies targeting tick control,
such as acaricide treatment and pasture management, are imperative for mitigating the impact of
ECF on cattle populations in regions like Karamoja where these ticks are prevalent (Okello et al.,
2013)
A study found a 36.4% prevalence of `A. variegatum` on goats in Nakapiripirit District,
demonstrating its potential for Cowdriosis transmission(Anya et al., 2019)

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 Namalu sub-county, Nakapiripirit District in northeastern Uganda lies within the Karamoja sub-
region, known for its semi-arid climate and extensive savanna grasslands. This ecological setting
forms a prime habitat for diverse tick species, posing a significant challenge to both livestock
and human health in the region. Here, we review the prevalence of ticks in Namalu sub-county,
Nakapiripirit District, highlighting key species and their associated diseases.

2.2.2 Dominant Tick Species:

`Rhipicephalus evertsi evertsi`: Commonly known as the brown ear tick, it readily infests cattle,
goats, and sheep. It is a primary vector of East Coast fever (ECF), a highly contagious and often
fatal disease affecting cattle with a prevalence rate of 7.8% (Etiang et al., 2024)
`Amblyomma variegatum`: This bont tick also targets cattle, sheep, and goats, transmitting
Cowdriosis, a rickettsial disease causing fever, weakness, and neurological symptoms with a
prevalence rate of 32.3%

2.3 Tick Resistance: A Global Threat with Regional and National Nuances
The emergence and spread of acaricide resistance in ticks pose a significant threat to animal
health and agricultural productivity worldwide. Controlling tick infestations and the transmission
of tick-borne diseases relies heavily on acaricides, but their effectiveness is increasingly
compromised by resistant tick populations. Understanding the geographical patterns and
mechanisms of resistance is crucial for implementing effective control strategies.(Production,
2022)

2.3.1 Global Trends:

Widespread and increasing: Acaricide resistance has been documented in ticks across all
continents affecting various species and acaricidal classes. The situation is particularly
worrisome for Rhipicephalus (Boophilus) microplus, the most economically important tick,
showing resistance to all major acaricidal classes in some regions(Githaka et al., 2022)
Multiple resistance mechanisms: Ticks employ different strategies to combat acaricides,
including target-site mutations, increased detoxification, and reduced acaricidal penetration.
Understanding these mechanisms helps develop resistance management strategies and prioritize
research efforts.
Economic and health impacts: Resistance contributes to increased tick burdens, reduced control
efficacy, and higher treatment costs for livestock farmers. Tick-borne diseases like babesiosis and
theileriosis can become more prevalent, impacting animal health and food security(Obaid et al.,
2022)

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2.3.2 Regional Trends:
Africa: Acaricide resistance is widespread in African countries, particularly against pyrethroids
and organophosphates. The threat to livestock production and livelihoods is significant(Githaka
et al., 2022)

2.3.3 National Trends:

Uganda: A recent study found that 93.5% of tick populations tested in Uganda had resistance to
at least one acaricide class. This highlights the critical need for evidence-based acaricide
resistance management strategies(Vudriko et al., 2016)
National studies:
Multi-acaricide resistance: 55.2% of resistant Rhipicephalus ticks in Uganda displayed
resistance to at least two acaricides (mainly pyrethroids and organophosphates)(Vudriko et al.,
2016)

2.3.4 Possible Causes of Resistance:

Overuse and misuse of acaricides: Frequent and inappropriate use of the same chemicals selects
for resistant tick populations.
Lack of acaricide rotation: Switching between different chemical classes is crucial to reduce
selection pressure.
Improper application: Incorrect doses, concentrations, or targeting can lead to incomplete tick
control and resistance development(Vudriko et al., 2016).

2.3.5 Potential Consequences:

Increased disease burden: Ticks act as vectors for diseases like East Coast Fever, with potentially
devastating consequences for livestock health and productivity.
Economic losses: Tick-borne diseases and acaricide resistance contribute to decreased milk
production, animal deaths, and higher treatment costs, impacting farmer livelihoods.
Food security: Reduced livestock productivity can threaten food security and livelihoods in
communities dependent on animal agriculture.

2.4 Plants as sources of pesticides/acaricides

Some of the plants reported in the present survey were previously validated for their anti-tick
activity through in vitro tests for example Azadirachta indica (Williams 1993) and Nicotina
tobacum (Choudhrey et al 2004)

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2.4.1 Ricinus communis
Riccinus communis is a native plant, commonly known as the castor oil plant, is a species of
flowering plant in the spurge family, Euphorbiaceae. Native to the Mediterranean region, Eastern
Africa, and India, has been explored for its potential as an acaricide, which is a substance used to
control or eliminate ticks and mites. A study by (Gosh et al.2013) showed a 95% acaricidal
mortality rate in a period of 14 days.

2.4.2 Ocimum basilicum.

Ocimum basilicum, commonly known as basil, has been investigated for its potential as a natural
acaricide and it showed an 80% mortality rate in ticks in a period of 15 days(Zhang et al. 2009)

2.4.3 Carica papaya

Papaya-based acaricides have been explored as alternative methods for tick and mite control in
agriculture and animal husbandry, offering potential benefits such as creating alternatives. The
pawpaw leaves extracts have also been used against hard ticks and have shown a potential
93.33% mortality rate(Ocloo et al., 2012)

2.4.4 Pyretthroids
Pyrethroids are a class of synthetic chemicals that are widely used as acaricides, which means
they are used to control and kill ticks and other arachnids like mites and spiders. Here's more
detail about pyrethroids:
Chemical Structure: Pyrethroids are derived from natural compounds called pyrethrins, which
are extracted from certain species of chrysanthemum flowers. They have a similar chemical
structure to pyrethrins but are more stable and have enhanced insecticidal properties(Hodoșan et
al., 2023)
Mode of Action: Pyrethroids work by targeting the nervous system of ticks and other insects.
They interfere with the normal function of nerve cell membranes by prolonging the opening of
sodium channels. This leads to hyperexcitation of the nervous system, causing paralysis and
eventual death of the tick.
Broad-Spectrum Activity: Pyrethroids have a broad spectrum of activity, meaning they are
effective against a wide range of tick species as well as other insects. This makes them valuable
tools for controlling tick infestations in agricultural, veterinary, and public health settings.
Persistence: Some pyrethroids have residual activity, meaning they can remain effective for an
extended period after application. This characteristic is particularly useful in controlling ticks in
outdoor environments where reinfestation is a concern(Shefali et al., 2021)
Formulations: Pyrethroids are available in various formulations, including sprays, dusts, and
impregnated materials (such as treated clothing or animal collars). These formulations allow for
different methods of application depending on the target pest and the environment.
Environmental Impact: While pyrethroids are generally considered safe for humans and other
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mammals when used according to label instructions, they can be toxic to fish and other aquatic
organisms. Care should be taken to avoid contamination of water bodies during
application(Hodoșan et al., 2023)
Resistance: Over time, some tick populations have developed resistance to pyrethroids due to
repeated exposure. Resistance management strategies, such as rotating different classes of
acaricides and using integrated pest management approaches, are important for maintaining the
effectiveness of pyrethroids and other tick control measures.

2.5 Acaricidal assays

Acaricidal assays are scientific experiments designed to evaluate the efficacy of substances or
formulations in killing or repelling ticks and mites. These assays typically involve exposing the
target arthropods to the test substances under controlled laboratory or field conditions and
measuring parameters such as mortality rates, repellency, or effects on reproduction and
development(Paramasivam, 2017)

2.5.1 Contact toxicity assays:

In these assays, ticks or mites are directly exposed to the test substance, either by applying it to
their bodies or by placing them in contact with treated surfaces. Mortality rates are recorded over
a specified period to assess the lethal effects of the substance(Eisen et al., 2018)

2.5.2 Fumigation assays:

These assays involve exposing the arthropods to vapors or aerosols of the test substance in a
closed environment. The mortality or knockdown effects of the fumigant on the ticks or mites are
then evaluated(Richter et al., n.d.)

2.5.3 Repellency assays:

Repellency assays assess the ability of a substance to deter ticks or mites from contacting or
feeding on a treated surface or host. Arthropods are given a choice between treated and untreated
surfaces or hosts, and their preference or avoidance behavior is observed and quantified(Luker,
2024)

2.5.4 Residual activity assays:

These assays assess the persistence of acaricidal activity over time by measuring mortality rates
or repellency effects at different intervals after treatment application. They provide information
on the longevity of the acaricidal effects of the test substance.

2.6 Life cycle and behavior of ticks

2.6.1 Life Cycle:

i. . Egg Stage: The life cycle begins with the female laying a batch of eggs. The number of
eggs varies among species, and environmental conditions influence their development.
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 ii. . Larval Stage: After hatching, larvae have six legs and seek a host for their first blood
meal. Once fed, they detach and molt into the nymphal stage.
iii. . Nymphal Stage: Nymphs have eight legs and, like larvae, seek another host for a blood
meal. After feeding, they detach, molt into the adult stage, and enter a period of
development known as the molting process.
iv. . Adult Stage: Adult ticks have eight legs and are sexually mature. Both male and female
adults seek hosts for blood meals. Mating often occurs on the host, and females engorge
with blood. After feeding, females drop off the host to lay eggs, completing the life
cycle(Apanaskevich & Oliver, 2014b).

2.6.2 Feeding Habits:

Obligate Blood Feeders: Ticks are obligate blood feeders at every stage of their life cycle. They
rely on blood meals for nutrition and reproduction.
Host Specificity: Different species of ticks exhibit host specificity, meaning they prefer specific
hosts. Some ticks have a broad host range, while others are more selective.
Feeding Duration: The duration of a tick's blood meal varies. It can range from several minutes
to several days, depending on the tick species, life stage, and host characteristics(Militzer et al.,
2021)

2.6.3 Behavior:
Questing: Ticks use a behavior called "questing" to find hosts. They climb vegetation, extending
their legs and waiting for a passing host. When a suitable host brushes against them, they grab
onto the host and start feeding.(Leal et al., 2020)
Chemotaxis: Ticks can detect hosts through chemical cues such as carbon dioxide, heat, and
specific odors emitted by hosts. This helps them locate and reach potential hosts efficiently.(Carr
& Salgado, 2019)
Attachment and Feeding: Ticks use specialized mouthparts to pierce the host's skin and feed on
blood. They secrete substances to prevent blood clotting and facilitate feeding. The feeding
process can transmit various pathogens if the tick is carrying them(Krcmar, 2012)

2.7 In vitro acaricidal testing

Through in vitro testing, this study aims to assess the efficacy of plant extracts or compounds
derived from selected medicinal plants in controlling tick populations. The process involves
selecting specific medicinal plants known for their pesticidal properties or traditional use in pest
control. These plants are then processed to extract active compounds, which are subsequently
tested against ticks in controlled laboratory conditions.
In these experiments, various parameters are evaluated, including the mortality rate of tick larvae
after exposure to plant extracts, the inhibition of tick growth or reproduction, and any adverse
11
effects on the ticks' physiology. By systematically investigating the acaricidal properties of
medicinal plants, researchers can identify promising candidates for further development as
natural alternatives to synthetic acaricides. Additionally, such research contributes to the growing
body of knowledge on botanical interventions for tick control, offering sustainable and
alternative solutions to combat tick-borne diseases and reduce or combat acaricide
resistance(Olivia chavez et al., 2023)

CHAPTER THREE: RESEARCH METHODS.

3.0 Methodology

3.1. Study area

Namalu sub-county in Nakapiripirit district is located in the northeastern part of the country in
the Karamoja subregion. Approximately 95 kilometers northeast of Mbale City the nearest large
city and 108 kilometers south of Moroto Town the largest urban centre in the Karamoja sub-
region and 320 kilometers northeast of Kampala, the capital and largest city of Uganda. Namalu
sub-county is located along the coordinates 1°45'39.6"N 34°36'28.8"E. Namalu sub-county has a
total of 5 parishes namely Lokatapan, Loperot, Kokuwam, Kaiku and grazing area with 16
villages
3.2 Study design

The study design incorporates both qualitative and quantitative elements to comprehensively
explore the efficacy of selected medicinal plants against ticks in Namalu sub-county,
Nakapiripirit District.

A sample of ticks will also be collected from kraals in Namalu sub-county by setting up baits or
hand-picking from the animals

Target species:

Collection of the correct plant species based on my prior selection and traditional knowledge
verification. Consulting with local communities or botanical experts for identification.

Plant parts: Collection of the specific plant parts traditionally used for tick control(e.g., leaves,
bark, whole plant).Aiming for sufficient quantities(e.g., 100-200g species)to allow for adequate
extract preparation.

Collection time: Choosing the optimal time for collection based on the plant's life cycle and
potential variation in active compound content to avoid collecting during periods of stress or
adverse weather conditions.

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Collection method: Employing appropriate methods to minimize damage to the plant and ensure
clean material. Wearing gloves and using sterile tools when necessary.

Sample processing and storage: Plant parts will be washed with clean water to remove debris and
soil. Dried thoroughly and shade-dried whenever possible to preserve active compounds and
stored.

3.3 Research Design:

The research design involves a mixed-methods approach, in data collection methods will include
survey and experiments to assess the acaricidal efficacy of medicinal plant extracts.

3.4 Research Approach:

The research adopts an exploratory approach to investigate traditional knowledge and practices
related to tick management, as well as empirical testing to evaluate the effectiveness of medicinal
plants as alternative acaricides. By combining qualitative insights with quantitative data on tick
mortality rates, the study aims to generate comprehensive findings to address the research
objectives.

3.5 Sample Size Selection:

The sample size for qualitative data collection, including interviews and community
consultations, will be determined through purposive sampling techniques to ensure
representation of diverse perspectives and knowledge holders within Namalu sub-county. Key
informants, including traditional healers, livestock farmers, and local experts, will be selected
based on their expertise and experience in traditional tick control methods.

For quantitative laboratory experiments, the sample size will be determined based on statistical
considerations to ensure adequate power to detect significant differences in tick mortality rates
between treatment groups. Sample size calculations will take into account factors such as effect
size, desired level of confidence, and expected variability in tick response to plant extracts.

3.6 Study Population:

The study population includes residents of Namalu sub-county, Nakapiripirit District,

particularly those involved in livestock rearing and traditional herbal medicine practices. Key
stakeholders such as traditional healers, farmers, and community leaders will be actively engaged
in the research process to facilitate knowledge sharing and collaboration.

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3.7 Identification of medicinal plants

With the use of indigenous knowledge on medicinal plants which could have earlier been used
by residents in Namalu sub-county, some plants are going to be selected

3.8 Transportation and Storage of Selected Plant Samples:

Transportation and storage procedures for selected plant samples are critical to preserve the
integrity and potency of the medicinal plant materials throughout the research process. The
following steps will be taken:

3.8.1 Transportation:

1. Collection Protocol: During plant collection, utmost care will be taken to avoid damage
to the plant material. Plants will be harvested using appropriate tools, and only the
required plant parts will be collected to minimize waste.

2. Packaging: After collection, the plant samples will be carefully wrapped in clean,
breathable material such as paper or cloth to prevent wilting or bruising during
transportation.

3. Storage Containers: Plant samples will be placed in sturdy, breathable containers such as
cardboard boxes or baskets to allow air circulation while protecting them from physical
damage.

4. Transport Vehicle: Plant samples will be transported in a dedicated vehicle equipped with
appropriate storage facilities to maintain optimal temperature and humidity conditions
during transit.

5. Avoidance of Contamination: Measures will be taken to prevent contamination of plant

samples during transportation, including avoiding contact with chemicals, pesticides, or
other potential contaminants.

3.8.2 Storage:

1. Cool, Dry Environment: Upon arrival at the research facility, plant samples will be stored
in a cool, dry environment away from direct sunlight to maintain their freshness and
potency.

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 2. Proper Ventilation: Adequate ventilation will be provided in the storage area to prevent
moisture buildup and fungal growth, which can compromise the quality of the plant
materials.

3. Labeling: Each plant sample will be clearly labeled with relevant information such as
species name, collection date, location, and collector's name to ensure traceability and
proper documentation.

4. Shelving or Racks: Plant samples will be stored on shelves or racks to facilitate

organization and prevent overcrowding, which can lead to damage or contamination.

5. Regular Inspection: Stored plant samples will be regularly inspected for signs of
deterioration, mold, or pest infestation. Any compromised samples will be promptly
discarded to prevent cross-contamination.

6. Security Measures: Access to the storage area will be restricted to authorized personnel
only to prevent unauthorized handling or tampering of plant samples.

3.9 Experiment design.

i. Materials

Gloves

Protective gloves made of latex shall be used for handling plant materials to avoid contamination
with pathogens

Petri dishes

Petri dishes, also known as petri plates or cell-culture dishes, are shallow, flat, cylindrical, or
round glass or plastic dishes with a loose-fitting lid. They are commonly used in laboratories for
culturing microorganisms and holding organisms.

Soxhlet extraction apparatus

A Soxhlet apparatus is a laboratory tool used for extracting compounds from a solid sample. It
consists of a round bottom flask, a condenser, a siphon tube, and a thimble. The solid sample is
placed in the thimble, which is then inserted into the round bottom flask. The solvent is heated in
the round bottom flask, causing it to evaporate and condense in the condenser. The condensed
solvent drips back into the round bottom flask, continuously extracting the compound from the
solid sample.

Extraction reagent. (methanol)

15
Methanol is a common extraction reagent used in various laboratory procedures, including the
extraction of organic compounds from solid or liquid samples. It is a polar solvent that is
miscible with water and many organic solvents, making it a versatile choice for the extraction of
medicinal plants.
Methanol will be used because of its ability to dissolve a wide range of organic compounds,
including polar and non-polar substances. It is also relatively non-toxic compared to other
solvents, making it a safer option for laboratory work. Additionally, methanol has a low boiling
point, which allows for easy evaporation after the extraction process.
Motor and pestle

For grinding the medicinal plant parts.

Measuring cylinders

Are calibrated glass or plastic containers. Will be used to measure the required amount of solvent
and extracts.

Weighing scale

For measuring sufficient quantities of the medicinal plant material to be extracted

Stop clock

For observing extraction time in which the ticks die after immersion into the extracts

ii. Extraction procedures

The plant materials prepared will be crushed to fine form to increase surface area for extraction
of active ingredients with suitable solvents using a Soxhlet apparatus.

Yield will be determined by; weight of extract (grams) X 100

Weight of plant material before extraction(grams)

The extracts will then be prepared for use.

iii. In vitro testing

In vitro tests will then be carried out by tick larvae immersion into the extracts and observed for
a period ranging from hours to days and the number of ticks that die will be established

16
Tick mortality will be calculated as; number of dead ticks x100

Total number of ticks(Shyma et al., 2015)

3.4. Data collection method and data collection tool

3.5. Data analysis.
Collected raw data will be stored in a Microsoft Excel database system for data management.
SPSS Windows will be used for data analysis. Mean tick mortality and related results of the
study will be expressed using descriptive statistics (mean ± standard error of mean, percentage,
and graph). One-way analysis of variance (ANOVA). All significant levels will be set at 95%
(Oliva Chávez et al., 2023)

3.10 Ethical Considerations:

Informed Consent: Prior informed consent will be obtained from all participants involved in
interviews, surveys, and experimental procedures. Participants will be provided with detailed
information about the research objectives, procedures, potential risks and benefits, and their right
to withdraw from the study at any time.
Confidentiality: All data collected, including personal information and research findings, will be
treated with strict confidentiality.
Respect for Cultural Norms: The research will respect and adhere to local cultural norms,
customs, and traditions. Sensitivity to cultural practices and beliefs related to medicinal plants,
livestock management, and community engagement will be maintained throughout the research
process.
Beneficence and Non-Maleficence: The research aims to contribute positively to the well-being
of local communities by generating knowledge that can enhance sustainable tick management
practices. Measures will be implemented to minimize any potential harm or risks to participants,
including adherence to laboratory safety protocols and ethical guidelines for plant collection and
experimentation
Research Integrity: The research will uphold principles of scientific integrity, transparency, and
rigor in all aspects of study design, data collection, analysis, and reporting. Any conflicts of
interest or biases will be disclosed, and research findings will be presented accurately and
objectively.

17
Questionnaire

Medicinal Plants Usage in Tick Control: Farmer Interview Questionnaire

Personal Information:
1. Name of the Farmer:
………………………………………………………………………………………………
…………….
2. Contact Information:
………………………………………………………………………………………………
…………….
3. Location:
………………………………………………………………………………………………
………………………………
4. Years of Farming Experience:
………………………………………………………………………………………………
Tick Infestation and Management:
5. Have you experienced tick infestations in your livestock? (Yes/No)
Yes

No

6. How do you currently manage tick infestations in your livestock?

Synthetic acaricides

Traditional methods (specify)

Others (specify)

7. Have you noticed any changes in tick populations or their resistance to conventional
acaricides over the years?

18
 Yes

No

If yes, please describe:

………………………………………………………………………………………………………
………………………………………………………………………………………………………
………………………………………………………………………………………………………
………………………………………………………………………………………………………
………………………………………………………
Knowledge and Use of Medicinal Plants:
8. Are you familiar with any medicinal plants used locally for controlling ticks?
Yes

No

If yes, please list the plants you are aware of:

…………………………………………………………………………………………………
…………………………………………………………………………………………………
…………………………………………………………………………………………………
…………………………………………………………………………………………………
…………………………………………………………………………………………………
…………………………………………………………………………………………………
…………………………………………………………………………………………………
…………………………………………………………………………………………………
…………………………………………………………………………………………………
……………………………………………………..
9. Have you personally used any medicinal plants for tick control on your farm?
● If yes, please specify the plant(s) and how you use
them:..........................................................................................................................
....................................................................................................................................
....................................................................................................................................
....................................................................................................................................
...................................................................................................................................

19
 10. How did you acquire knowledge about the use of these medicinal plants? (e.g., family
tradition, community knowledge, personal
experimentation)....................................................................................................................
..............................................................................................................................................
11. Can you describe the preparation and application process for using medicinal plants
against
ticks?......................................................................................................................................
................................................................................................................................................
................................................................................................................................................
................................................................................................................................................
..............................................................................................................................................
12. Have you observed any effectiveness in using medicinal plants compared to synthetic
acaricides?
Yes

No

If yes, please describe your observations:

…………………………………………………………………………………………
…………………………………………………………………………………………
…………………………………………………………………………………………
…………………………………………………………………………………………
…………………………………………………………………………………………
…………………………………………………………………………………………
…………………………………………………………………………………………
………………………………………………………………
Challenges and Opportunities:
13. What challenges do you face in using medicinal plants for tick
control?..............................................................................................................................................
............................................................................................................................................................
............................................................................................................................................................
............................................................................................................................................................
............................................................................................................................................................
............................................................................................................................................................
......................................................................
14. Do you see any potential benefits or opportunities in further exploring the use of
medicinal plants for tick
management?.........................................................................................................................
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 ................................................................................................................................................
................................................................................................................................................
................................................................................................................................................
................................................................................................................................................
....................................................
15. Are there any specific types of support or resources that would help you utilize medicinal
plants more effectively for tick
control?..................................................................................................................................
................................................................................................................................................
................................................................................................................................................
................................................................................................................................................
........................................
General Information: 16. Do you have any additional comments or insights you would like to
share regarding tick control or the use of medicinal plants in
agriculture?........................................................................................................................................
............................................................................................................................................................
............................................................................................................................................................
............................................................................................................................................................
...................................................
Thank you for your participation! Your input is valuable for our research on sustainable
tick management practices.

21
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