NATIONAL RESEARCH FUND PROPOSAL

Principal Researcher / Investigator

Name: DR GRACE O. ADEOYE

Institution: UNIVERSITY OF LAGOS, NIGERIA

Department: Department of Zoology, University of Lagos, Akoka, Yaba, Lagos, Nigeria.

Telephone No: 08035373776

E-mail Address: goadeoye@gmail.com, gadeoye@unilag.edu.ng

Co-Researcher (s) (if any):

1. Name: DR MONSURU A. ADELEKE (Dept. of Biological, Osun State University)

2. Name: DR CHRISTIANAH AYOLABI (Dept. of Microbiology, University of
Lagos)

Project Title: PREVALENCE AND MOLECULAR EPIDEMIOLOGY OF URINARY

SCHISTOSOMIASIS, SOIL-TRANSMITTED AND ROTAVIRAL INFECTIONS IN PRE-
SCHOOL AGED CHILDREN IN OSUN STATE, NIGERIA

Executive Summary: Schistosomiasis, soil-transmitted and rotaviral infections are among the
most prevalent afflictions of humans who live in areas of poverty in the developing world,
including Nigeria. There are many reports of work done on the prevalence of schistosomiasis and
soil-transmitted helminth infections in School-aged children in different States of Nigeria.
However, very little work has been carried out on pre-school-aged children. The pre-school-aged
children are not included in the treatment plan in the country. Hence, the need for this study,
which aims at determining the prevalence and intensity of urinary schistosomiasis and soil
transmitted and viral infections. The factors responsible for the transmission of the diseases
would be examined. Urine and stool samples of 1,800 pupils selected randomly would be
collected and screened for the different infections. The haemstick / Dipstick (Combi-9), urine
filtration technique, Kato-Katz kit, ELISA and PCR techniques would be employed in screening
the urine and stool samples. Results obtained will help in determining application of Mass Drug
Administration among the Pre-school aged children or not. The findings from this study may
also lead to future intervention of the diseases at the pre-school level in Osun State. The study
may also be replicated in the other States of the Federation.

Introduction:

Schistosomiasis, soil-transmitted and rotaviral infections are among the most prevalent

afflictions of humans who live in areas of poverty in the developing world. Schistosomiasis is a

1
disease caused by the parasitic trematode belonging to the genus Schistosoma and it is

transmitted by the freshwater snail intermediate hosts (1). The disease is one of the leading

causes of social morbidity in the tropical region affecting over 200 million people worldwide (2).

Schistosomiasis is predominantly found in communities whose residents usually have contact

with freshwater habitats either due to the agricultural, domestic and recreational needs or due to

lack of reliable water for drinking, washing and bathing (3 -9).

The occurrence of soil-transmitted helminthiasis (STH) is associated with socio-economic,

environmental and other factors like ignorance of simple health promoting factors and

overcrowding, limited access to clean water, tropical climate and low altitude (10). School-aged

children are one of the groups at high risk for intestinal parasitic infections. Factors like poor

development of hygienic habits, immune system and over-crowding contributes to infection (11).

Rotaviral infection in school children leads to diarrhoea. Diarrhoea is a major cause of paediatric

morbidity and mortality, causing over 5 million deaths per annum (12 & 13) especially in

developing countries where malnutrition is common (14). The World Health Organization started

the diarrhoeal disease control programme in 1980 with the objective of decreasing diarrhoea

illness and death among children in developing countries. The Federal Government of Nigeria

also promoted enlightenment programmes for the prevention of diarrhoea. In spite of these

control measures, diarrhoea still remains the second main cause of death among children under 5

years in the developing world including Nigeria. Viruses account for 75% of infantile

gastroenteritis and are known to be potent inducers of diarrhoea, vomiting and subsequent

dehydration. Rotavirus (RV) is the most severe viral pathogen in paediatric diarrhoea (15 & 16).

2
 Lot of studies have been done on school-aged children (3-9). It is therefore necessary to

examine the pre-school aged children in Osun State for urinary schistosomiasis, soil-transmitted

and rotaviral infections.

Problem Statement / Justification:

The creation of dams to carter for irrigational activities in Nigeria has been accompanied with

many public health problems, especially the spread of schistosomiasis (17 & 18). Dams have

been known to provide conducive habitats for snail intermediate hosts which normally translate

to intense disease transmission in communities around the dams (19 – 21). In addition, all school

children are exposed to soil-transmitted infections through various activities carried out at school

especially during play and sporting activities. Polyparasitism in school children has been

reported by Hotez et al. (22) and Adeoye et al. (9). In many paediatric clinics in Nigeria, routine

rotavirus screening is not performed probably due to its clinical spectrum of signs and symptoms

similar to gastroenteritis caused by other pathogens. As a result, the actual burden of rotavirus

diarrhoea among children less than 5 years of age is underestimated. Definitive diagnosis of

rotavirus is fundamental to the treatment and management of infected children and also to the

prevention of infection. Moreover, the rotavirus strains prevalent in Nigeria have not been well

characterized and not much work has been done on viral infections in Osun State.

Annual Mass Drug Administration (MDA) of Praziquantel and Albendazole for schistosomiasis

and soil-transmitted helminthiasis has been prescribed for school-aged children in endemic areas

However, the reports of high intensity of urinary schistosomiasis among infants (7 & 21) call for

renewed interest on the need to understand the role of the infantry on the epidemiology of

urinary schistosomiasis, soil-transmitted infections and rotoviral gastroenteritis. Efforts should

3
be made to curtail the diseases among the pre-school age groups who are excluded in the

treatment regime by the WHO protocols.

Objective(s) of the Study:

1. To determine the prevalence and intensity of urinary schistosomiasis and soil-transmitted

infections among the Pre-school aged children in Osun State.

2. To determine the prevalence of rotavirus gastroenteritis among the pre-school-aged

children in Osun State.
3. To determine the rotavirus genotypes circulating among under 6 years children

presenting with diarrhea.

4. To examine the level of co-infections among the pupils.

5. To determine the risk factors for the infections among the pre-school children

6. To compare the level of prevalence of infection by parasitological techniques with those

of Enzyme-linked Immunosorbent Assay (ELISA) and Polymerase Chain Reaction

(PCR).

Literature Review:

Soil-transmitted Helminthiasis (STH) constitutes great percentage of Neglected Tropical

Diseases (NTD). Approximately 85% of the neglected tropical disease (NTD) burden results

from helminth infections; hookworm is the most common Soil-Transmitted Helminthes (STH)

and the most common NTD in Sub-Sahara Africa (SSA) (23). Approximately one-third of the

world’s hookworm today occurs in SSA (24), with the greatest number of cases occurring in

Nigeria (38 million) and the Democratic Republic of Congo (31 million). Schistosomiasis is the

4
second most prevalent NTD after hookworm. Approximately 207 million persons are infected

with schistosomiasis worldwide (25) which leads to the loss of approximately 1.53 million

disability-adjusted life years (26). Of the world’s 207 million estimated cases of schistosomiasis,

93% occur in SSA (192 million), with the largest number in Nigeria (29 million) (27). The four

most common STHs are round worms (Ascaris lumbricoides), whipworms (Trichuris trichiura),

and the antropophilic hookworms (Necator americanus and Ancylostoma duodenale). The

greatest numbers of STH infections occur in the Americas, China and East Asia, and Sub-

Saharan Africa (28 & 29). It is estimated that 173 million and 162 million people are infected in

SSA with Ascaris and Trichuris, respectively, with 36 million school-aged children infected with

ascariasis and 44 million with trichuriasis (30). For both infections, the largest number of cases

occurs in Nigeria, where co-infections with hookworms are common (31). In children, these

parasitic infections can have adverse effects on physical growth and cognitive development (10).

Jardiam-Batelho (32) and Sakti (33) suggest that children with multiple parasitic infections tend

to experience more severe health problems than those with only one infection. Mazigo (34)

observed co-infection of schistosomiasis and hookworm and S. mansoni and Plasmodium

falciparum among children in Tanzania. Intervention against soil-transmitted helminths (STH)

and Schistosoma infections are based on regular anti-helminthic treatment, improved water

supply, sanitation and health education (35 & 36).

Methodology (Should include description of study area/site/subjects, data collection and

data analysis):

Study Area / Site:

5
The study will be carried out in Osun State of Nigeria. Osun State is an inland State in south-

western Nigeria. Its capital is Osogbo. It is bounded in the north by Kwara State, in the east

partly by Ekiti State and partly by Ondo State, in the south by Ogun State and in the west by Oyo

State. Osun State is organized into 30 Local Government Areas; geopolitically stratified into

Three (3) Senatorial Districts (Figure 1). 

Figure 1: Map of Osun State showing the Senatorial districts

Data Collection from Study Participants:

6
Ethical Clearance

Ethical clearance and approval will be obtained from the Ethical Committee of Osun State

University, Osogbo, Nigeria. After the receipt of the ethical clearance, advocacy visits would be

made to the Osun State Ministries of Health and Education. Prior to the commencement of the

study, permission would be sought from the various Local Government Authorities that would be

involved in the study. Informed consent will be sought from the parents and care-givers of the

participants to be enrolled for the study. The parents and care-givers of the pre-school children

(less or equal to 6 years old) will be mobilized for the study.

Data Collection:

Sample Size Determination

The sample size for this study would be calculated using single proportion formula at 95%
Confidence Interval (CI) level (Z(1-1/2a)=1.96), an expected prevalence of 50% since there was
no study conducted regarding this topic in the area and 5% marginal error. Then, the sample size
will be calculated as n=((Z(1-1/2a) 2P(1-P)/d2, where n=sample size, P=proportion of problem in
the study area, (Z(1-1/2a)= CI of 95%, d = Marginal error to be tolerated. By adding 10%
contingency, 1537 pupils (minimum number) would be included in our study. This number
would be rounded up to 1800 pupils for the study.

Experimental Design

. The study will utilize cross-sectional multi-sampling techniques. Twelve Nursery /Primary

schools will be randomly selected in each senatorial district (consisting Six Government owned
Public schools and Six Private Owned Nursery/ Primary Schools). Fifty pupils will then be
randomly selected among the pupils in each selected school. The random sampling will be done
by balloting. A total of 1,800 pupils would be sampled from the three Senatorial Districts.

Demographic data including the age and sex of the enrolled pre-school children (1 to 6 years old)
whose parents/ care-givers have consented to participate in the study will be recorded in the field
book and sample numbers would be assigned to them.

Assessment of Transmission Risk Factors

The height and weight of each child will be taken with cloth tape and weighing scale. Baseline
information on these diseases will also be obtained from the community members using

7
structured questionnaires. The survey will also assess other facilitating factors that aid the
transmission of Schistosomiasis, Soil transmitted helminth infections and viral gastroenteritis in
Osun State. There would be Focus Group Discussion with the Parents, Care Givers and Teachers
of the Pupils as to the activities of the children in and out of the home, especially at school.

Data Collection in the Schools

Data collection in the Schools is a comprehensive team effort, which requires thorough
preparation and excellent coordination. It is extremely important for the survey team to inform
school authorities, school children, community leaders and parents, of the specimen collection
date well ahead of time.

Mobilization/Sensitization
The State Universal Basic Education Board (SUBEB) Personnel will be used in the mobilization
of the schools. Their involvement will also help to maximize school attendance on the day of the
survey and minimize non-respondents.

Survey team
There will be one survey team comprising of laboratory scientists, technologists, Survey
recorders, field guides, and drivers. There shall be a team leader for coordination of the team.
The team composition and team member responsibilities are shown in the table below:

Table 1: Survey team members and their responsibilities

Role Suggested numbers Responsibilities
Survey Recorder 2 The recorder has the overall responsibility in
organising and overseeing the epidemiological
information collection activities, and therefore
should have a thorough knowledge of what to be
collected and how they are collected. The recorder
is also responsible for completing all reporting
forms including the lab results.
Laboratory 2 Collection of stool and urine samples, correctly
technologist labelling and packaging of samples, testing for
haematuria, and preparation of Kato-Katz and
filtration slides. Preparation for ELISA & PCR
techniques
Field guide 2 Notification, mobilization of the selected schools
and organizing the students
Driver 2 Safe-keeping and care of vehicles and other
assigned duties.
Laboratory 2 Carry out microscopic examination of all prepared
scientist slides; assist in running the ELISA and PCR.
SMOH Program 1 For advocacy, sensitization & mobilization at the
Co-ordinator State Level
SUBEB 1 For advocacy, sensitization & mobilization at the
School Level
LGA Programme 1 For advocacy, sensitization & mobilization at the

8
Co-ordinator LGA and Community Levels

Training of Team Members, Monitoring and Supervision of the survey exercise will be carried
out by the Principal Investigator and the two Researchers.

Collection of Urine and Faecal Samples from the Pupils

Urine and faecal samples would be collected from the pupils between the hours of 9.00 to

11.00am. Two (2) capped sterile specimen bottles will be labelled with specific identification

(ID) numbers for each pupil, one for urine and the other for stool collection. The container for

the stool has wooden applicator stick. All samples collected would be examined the same day.

Screening of Urine and Faecal Samples for Infection

For Schistosoma haematobium

During this survey four screening techniques will be used for S. haematobium:
 Haemstick for detection of micro-haematuria
 Syringe Urine filtration for S. haematobium eggs (determination of intensity)
 Enzyme-linked Immunosorbent Assay (ELISA)
 Polymerase Chain Reaction (PCR)

For Schistosoma mansoni and Soil Transmitted Helminths (STH) Infections

 Kato/Katz method will be used to determine prevalence and intensity of worms (eggs per
gram of faeces – epg).

For Rotaviral Antigen

 The stool samples would be screened for the presence of rotavirus antigen using Enzyme-
linked Immunosorbent Assay (ELISA)
 Rotavirus-positive samples will be characterized molecularly by Reverse-Transcriptase
Polymerase Chain Reaction (RT-PCR).

Data Analysis

The data will be analyzed with Chi-square and Student t-tests using SPSS version 16 (SPSS Inc,

Chicago, Illinois) and Epidata statistical software. The two statistical tools will be used to

compare the prevalence, intensity, water contact activities, disease perception and transmission

risk factors in the study areas.

9
Results (Expected Outputs / Results)

The prevalence, intensity and source of infection of the pre-school children in relation to urinary

schistosomiasis, soil-transmitted infections and rotaviral gastroenteritis will be known in the

study area. This will help to justify the need or otherwise for the inclusion of pre-school children

in the Mass Drug Administration or treatment policy of the Ministry of Health. A follow-up

study will focus on Intervention for these diseases at the Pre-school level as oral fomulation (oral

Praziquantel) for children are being developed for schistosomiasis and soil-transmitted

helminthiasis.

Work Plan / Time Frame

The proposed time frame for this study is 18 months. The Work Plan is shown below:

S/ TASK WHO JUL – OCT – JAN – APR – JUL – OCT –

N SEP DEC MAR JUNE SEP DEC

2015 2015 2016 2016 2016 2016

1. Ethical Principal 

Clearance & Investigator

Advocacy, (PI) & the 2

Mobilization Researchers

& (Research

Sensitization Team)

2. Selection of Research & 

Survey Survey

Sites /Schools

10
 Informed Teams

Consent,

Training of

Survey Team

& Data

Collection in

Senatorial

District (SD)

3 Selection of Research & 

Survey Survey

Schools, Teams

Informed

Consent,

Training of

Survey Team

& Data

Collection in

SD 2

4. Selection of Research & 

Survey Survey

Schools, Teams

Informed

11
 Consent,

Training of

Survey Team

& Data

Collection in

SD 3

5. Collation and Research 

Analysis of Team

Data,

Report

Writing

6. Presentation Research 

of Reports at Team

the LGA and

State Levels,

Presentation

of Reports

and Findings

at the

National &

International

Conferences

12
Budget:

S/N DESCRIPTION OF CATEGORY & NO OF COST PER TOTAL

ITEM / NO. OF PEOPLE TRIPS & TRIP / DAY COST

ACTIVITY/ DAYS (NAIRA) (NAIRA)

EQUIPMENT

1. Ethical Clearance & RESEARCH 3 (2 days N15,000 X 2 N270,000

Advocacy, TEAM (3) each per X 3 X 3

Mobilization & trip)

Sensitization

2. Selection of Survey RESEARCH 2 Trips (3 N15,000 x 3 N270,000

Sites /Schools TEAM (3) days per x 2 x3

Informed Consent, trip)

Training of Survey SURVEY TEAMS

Team & Data (13): 1 Trip (5

N15,000 x 5 N150,000
Collection in LabScientist (2) Days)
x1x2
Senatorial District
Lab.Technicians (2) N15,000 x 5 N150,000
(SD) 1
Recorders (2) x1x2
N100,000
N10,000 x 5

Field Guide (2) x1 x2

N20,000
N2,000 x 5 x

Drivers (2) 1x2

N30,000
N3,000 x 5 x
SMOH Prog. Co- 1x2

13
 ord.(1) N10,000 x 5 N100,000

SUBEB (1) x1x2

N10,000 x 5 N100,000

LGA Co-ord. (1) x1x2

N2,000 x 5 x N20,000

1x2

3. Selection of Survey RESEARCH 2 Trips (3 N15,000 x 3 N270,000

Sites /Schools TEAM (3) days per x 2 x3

Informed Consent, trip)

Training of Survey SURVEY TEAMS

Team & Data (13): 1 Trip (5

N15,000 x 5 N150,000
Collection in LabScientist (2) Days)
x1x2
Senatorial District
Lab.Technicians (2) N15,000 x 5 N150,000
(SD) 2
Recorders (2) x1x2
N100,000
N10,000 x 5

Field Guide (2) x1 x2

N20,000
N2,000 x 5 x

Drivers (2) 1x2

N30,000
N3,000 x 5 x
SMOH Prog. Co- 1x2
N100,000
ord.(1) N10,000 x 5
SUBEB (1) x1x2
N100,000
N10,000 x 5

14
 x1x2

LGA Co-ord. (1) N2,000 x 5 x N20,000

1x2

4. Selection of Survey RESEARCH 2 Trips (3 N15,000 x 3 N270,000

Sites /Schools TEAM (3) days per x 2 x3

Informed Consent, trip)

Training of Survey SURVEY TEAMS

Team & Data (13): 1 Trip (5

N15,000 x 5 N150,000
Collection in LabScientist (2) Days)
x1x2
Senatorial District
Lab.Technicians (2) N15,000 x 5 N150,000
(SD) 3
Recorders (2) x1x2
N100,000
N10,000 x 5

Field Guide (2) x1 x2

N20,000
N2,000 x 5 x

Drivers (2) 1x2

N30,000
N3,000 x 5 x
SMOH Prog. Co- 1x2
N100,000
ord.(1) N10,000 x 5
SUBEB (1) x1x2
N100,000
N10,000 x 5
LGA Co-ord. (1)
x1x2
N20,000

15
 N2,000 x 5 x

1x2

5. TRAINING OF 25 (Twenty-five) 2 Days N50,000 per N150,000

SURVEY TEAM PARTICIPANTS Training Venue /

(VENUE & Per Training /

REFRESHMENT) Senatorial SD (3 SD)

N150,000
District (3 N1000 /

SD) person / day

x2

6. Collation and RESEARCH Two Trips N15,000 / N360,000

Analysis of Data, TEAM (3) of 4 days day / person

Report Writing Each x2x4x3

7. Presentation of RESEARCH 3 trips of 2 N15,000 x 2 N270,000

Reports at the LGA TEAM (3) Days each x 3 x 3

and State Levels, at the 3

Presentation of SDs

Reports and Findings RESEARCH 1 N100,000 x 3 N300,000

at the National & TEAM (3) at Conference

International National Conf. Each

RESEARCH N600,000 x 3 N1,800,000
Conferences 1
TEAM (3) at Conference

International Conf. Each

16
8. EQUIPMENT Haemstick for N10,000

detection of

haematuria

Urine Filtration Kit N200,000

for S. haematobium

Kato Katz Kit for N300,000

faecal Screening

ELISA Machine & N2,000,000

Washer

PCR Machine N2,000,000

Electrophoresis N400,000

Tank & set up

Photodocumentatio N1,500,000

n Chamber

Centrifuge (Mini & N400,000

ordinary)

Heating Block N300,000

Vortex machine N400,000

Microwave N30,000

PCR kits N3,000,000

&Reagents

17
 ELISA kits & N3,000,000

Reagents

9. CHEMICALS & N10,000,000

CONSUMABLES

10. TRANSPORT, N5,000,000

VEHICLE HIRE &

FUEL

10. MISCELLENOUS N3,466,000

(10%)

11. UNILAG (5%) N1,733,000

12. TETFUND (5%) N1,733,000

13. TOTAL N41,592,000

The total budget is Forty-one Million, five-hundred and ninety-two thousand naira

(N41,592,000).

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