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Emergence of Sh 22.6 kDa Antigen Among the
Schistosoma haematobium Infected People in
Riverine Communities of the Extreme North Western
Nigeria
Suleman Jafaru
Sokoto State University https://orcid.org/0000-0001-9486-0797
Isyaku Nike Tawakaltu
Kebbi State University of Science and Technology
Ukatu Victoria Ebere
Kebbi State University of Science and Technology
Bagudo Ahmad Ibrahim
Kebbi State University of Science and Technology
Research Article
Keywords: Antigen, Emergence, Filtration Technique, PCR, Riverine Communities, Schistosoma
haematobium, SDS-PAGE, Urine
Posted Date: August 30th, 2024
DOI: https://doi.org/10.21203/rs.3.rs-4843645/v1
License: This work is licensed under a Creative Commons Attribution 4.0 International License.
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Abstract
Background
A cross-sectional study was conducted to investigate the emergence of Sh 22.6 kDa antigen among the
S. haematobium
infected people in riverine communities of the Northwestern part of Nigeria because
urinary schistosomiasis is one of the most pathogenic neglected tropical diseases that emerge and
reemerge-in-Nigeria. Nine hundred (900) urine samples were collected and analysed using the standard
ltration method, all microscopy-positive samples were subjected to PCR analysis for conrmation of the
positivity, and the PCR-positive samples were subjected to SDS-PAGE and western blotting analysis to
investigate the presence of Sh 22.6 kDa.
Results
Out of 276 PCR-positive individuals, 6.2% (17/276) were harbouring Sh 22.6 kDa antigen. The distribution
of the antigen was signicantly associated with the location (P > 0.05); however, a higher frequency was
observed in Shagari [7.1% (6/84)]. Males were more highly infected with the antigen [7.1% (14/197)] than
females [3.8% (3/79)] and there was a signicant association (P = 0.000). Although, the age group is not
signicantly associated with the antigen frequency (P > 0.05), infected people aged ≥ 56 years old had
the highest Sh 22.6 kDa antigen [40.0% (2/5)] than other age groups. Bloody urine experience (P = 0.017)
and urine colour (P = 0.000) were the study area's clinical manifestations associated with Sh 22.6 kDa
distribution.
Conclusion
The present study showed that a small proportion of the infected people harboured
S. haematobium
which has Sh 22.6 kDa antigen. Therefore, health education campaigns and mass chemotherapy should
be considered by the authorities concerned in the ght against urinary schistosomiasis in the area.
Background
The most dangerous species of schistosomes in Nigeria is
S. haematobium
, which can lead to severe
renal complications affecting cognitive domain and urinary tract pathology (Santos et al., 2021). Apart
from pathology, there is evidence that
S. haematobium
can affect nutritional status and lower levels of
haemoglobin (Hb) by decreasing food intake and increasing blood loss, which can result in iron
deciency anaemia (Atalabi et al., 2016). The World Health Organization recommended that the National
Schistosomiasis Control Programme be implemented in Nigeria in 1988 to provide regular anti-
helminthic treatment to at least 75% of school-age children in endemic areas of Nigeria (Daumerie et al.,
2010), However, the disease is still emerging and reemerging in the nation (Ladan et al., 2012;
Mohammed et al., 2020; Jiya et al., 2022).
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Antigens are soluble materials created by parasites or chemicals found on their body surface; the
majority of antigens are proteins, but some are polysaccharides from the parasites' bodies (Sepulveda et
al., 2010). Antigens typically consist of a large number of determinants that may repeat molecular
structures or be distinct from one another (Sepulveda et al., 2010; Casacuberta-Partal et al., 2021;
Suleiman et al., 2022). The term "resistance antigens" refers to any proteins, toxins, or other chemicals
that the parasites produce to withstand the effects of an antiparasitic drug to which they were previously
susceptible (Vale et al., 2017). It is problematic when a parasite population's resistance antigen
percentage rises because it gives the parasites the ability to withstand treatment with an antiparasitic
drug that has previously proven to be effective (Black et al., 2010). Due to its greater capacity to quickly
produce various resistant antigens on the surface of its teguments,
S. haematobium
induces varying
degrees of resistance (El-Kady et al., 2020).
An antigen called 22.6 kilo Dalton is a resistant antigen of Schistosomes belonging to the Cathepsin B
family which circulates in the infected individuals and is present in both adult worms and schistosomula,
this antigen confers immunity to the parasite against the host's immunity (Asuming-Brempong et al.,
2022). Numerous global investigations examined the existence and roles of 22.6 kDa antigen in
Schistosoma species
(Markakpo et al., 2015). It was reported that in
S. mansoni
, human IgE response
against 22·6 kDa antigen known as Sm 22·6 kDa (Fitzsimmons et al., 2004). Research in Kenya indicated
that IgE to Sm 22·6 may contribute to resistance or be a marker of resistant parasites (Galazka, 1993;
Mwai et al., 2016; Pearson et al., 2021). Another research on
S. japonicum
(Sj 22·6 kDa) among the
infected communities in the Philippines showed that sj 22.6 kDa is a major target for the human IgE
response (Santiago et al., 2018). Also, another on
S. haematobium
antigen (Sh 22·6 kDa) from Gabon
showed that after receiving treatment 35.5% of the infected individuals who were not completely cured
were harboring Sh 22.6 kDa antigen (Fitzsimmons, 2005). Praziquantel which is the drug mostly used for
the treatment of schistosomiasis in Sokoto was found with resistant and reduced effectiveness against
the parasites in some Countries such as Brazil (Da Silva
et al.
, 2005), Kwale district of Kenya (Vinkeles et
al., 2014), Senegal (Senghor et al., 2015) and Egypt (Di Bella et al., 2018); also, the resistance may likely
arise when the parasites produce the resistant genes or antigen (Pinto-Almeida et al., 2016).
S. haematobium
was the most re-emergent species among the
Schistosoma species
in the riverine
communities of Sokoto (Singh et al., 2016). Despite its record as the most pathogenic species of
schistosomes in Nigeria (El-Kady et al., 2020). However, the reports on whether the antigens were among
the factors increasing the reoccurrence in the area were scanty. Therefore, the identication of Sh 22.6
kDa antigens is very important by considering its advantages in identifying whether it is among the
factors contributing to the persistence of the parasite despite the mass chemotherapy of the individuals
in the areas. Also, it is valuable in assessing the ecacy of vaccine programs that can be expected in the
future when anti-parasite vaccination becomes available. It is hoped that present studies provide reliable
data which serve as a guide for the implementation of effective preventive and control measures in the
study area and other endemic parts of the country.
Methods
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Study Area
Riverine communities of Sokoto State were the study areas for the present study, Sokoto is one of the
Nigerian states found in the extreme northwest of the country, the state has a land area of 28,232.37
square kilometres with a coordinate of 13° 0' 21.1428'' N and 5° 14' 51.1872'' E, bordered with the
Republic of the Niger to the east, Kebbi State to the west and north, and Zamfara State to the south.
Sokoto is the state's capital and largest city (Topographic Sheet, 2010), and also has an estimated
population of more than 4.2 million (NPC, 2019). Sokoto State has an average temperature of 28.3°C
(82.9 °F). Although the area is generally very hot, the dryness helps keep the highest temperatures below
40°C (104.0°F) for most of the year. The warmest months are February to April when daytime
temperatures can exceed 45°C (113.0°F) (Kasim and Usman, 2021). The harmattan wind blows Sahara
dust over the region and dominates the climate from June to October which is the rainy season; this
wind reduces sunlight, which lowers temperatures and causes dust to accumulate in homes (Jibrillah
et
al.,
2019).
Hausa/Fulani people make up the majority of the population in Sokoto state, but there are also Yoruba,
Igbo, Kabawa, Zamfarawa, and Zabarmawa other tribes like Dakkarawa, Igala, Ibra, Nufawa, and
Gwarawa also live there (Arshad and Odeh, 2014).People are highly dependent on agricultural activities
such as farming and rearing animals during rainy and dry seasons, the people are highly dependent on
both subsistence and commercial farming that consists of growing crops from the oodplains of the
Sokoto-Rima River system, the state is also covered with richalluvial soil which allow the cultivation of
many cereals including rice and millet, guinea corn, maize, wheat and beans. During the dry season, the
people of Sokoto practice irrigation farming that allows the cultivation of tomatoes, cabbages, carrots,
cucumbers, onions, ginger, and many other vegetables (Ismail and Oke, 2012).
Study Population
The study population for the present research consisted of
S. haematobium
-infected people aged ve (5)
years and above residing in the riverine communities of Sokoto State.
Research Design
A cross-sectional research design involving a simple random sampling technique was employed in the
present research. Meanwhile, out of the identied Local Governments with riverine communities, four (4)
Local Governments (Wamakko, Kware, Goronyo, and Shagari) were assigned as sample collection
centres based on convenience.In each selected centre members were gathered either at the palace of
the District Head, outreached at their occupational places, or in their residences with the help of the
community focal person, and were selected using a simple random selection, similarly, the aim, the
objectives, the signicance, the rules and the regulations of the studies were explained to the
participants using local language (Hausa). A clean universal specimen bottle with a screw cap was given
to each participant to provide urine samples between 9:00 to 11:30 am, each sample was tagged with a
serial number correspondent to the serial number on the questionnaire that carries information about the
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participant, the samples were subjected to physical observations, and then analyzed in the laboratory
(Agi and Okafor, 2005).
Questionnaire Administration
A semi-structured questionnaire was designed and administered to each of the participants. The
questionnaire consisted of three (3) sections namely: Demographic information, clinical manifestations
and results sections (Asmaa
et al.,
2020).
Data Collection
Qualitative and quantitative data were collected from the eld and laboratory during the present study.
The qualitative data collected were place of living, gender (males and females), presence of
S.
haematobium
(Yes or No), dicult urination experience (Yes or No), painful urination experience (Yes or
No), urine colour (light yellow, dark brown, light brown or blood brown), result from com B 11 strip
(normal, proteinuria or haematuria), and presence of Sh 22.6 kDa (Yes or No). The collected quantitative
data was the intensity (number of
S. haematobium
eggs per 10ml of urine sample) and age of the
participants.
Collection of Urine Sample
Fifty millilitres (50 ml) screwed cap free grease plastic bottle was offered to each participant to collect
his/her middle to the terminal end urine sample from 10:30 to 11:30 am, a serial number was assigned
to each sample bottle correspondent to the serial number of the participant’s questionnaire. Each
collected sample was maintained at 40C and transported to the parasitology laboratory, Department of
Zoology, Usmanu Danfodiyo University, Sokoto for the analysis (A
et al.,
2016).
Filtrationof the Collected Urine Samples and Microscopy
Each sample was ltered using the standard method recommended by WHO (2002). The sample was
ltered through Whatman lter paper 0.5mm using a ltration machine within ve minutes, each lter
paper was transferred onto the carbon paper, and three drops of ninhydrin and two drops of Logu’s
iodine solutions were added respectively on the contents of the lter paper, the paper was allowed to dry
at room temperature and observed microscopically using X10 the X40 objective lens.
DNA Extraction
Deoxyribose Nucleic Acid (DNA) was extracted from each urine sample tested positive using QIamp DNA
mini kit (QIAGEN, Hilden, Germany), form each urine sample two hundred microliter (200 μl) was placed
into a DNA extraction tube (Microtube), 300 μl of lysis buffer was added into the tube, another 500 μl of
absolute ethanol was added into the same tube and incubated for the period of 10 minutes, the solution
was transferred into spine column and centrifuged at 8,000 rpm for 1 minute, the supernatant was
discarded and the process was repeated for the second time, subsequently, 500 μl of wash buffer (70%
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ethanol) was added to the sediment and centrifuged at 8,000 rpm for 2 mins, the supernatant was
discarded and the process was repeated, another 500 μl of wash buffer was added into the solution and
centrifuged at 13,000 rpm for three minutes, the supernatant was discarded, the solution was transferred
into the microtube (Eppendorf tube), 50 μl of molecular grade water was added and centrifuged at
13,000 rpm for 1 minute, the supernatant was discarded and nally, the extracted DNA was stored for the
polymerase chain reaction (PCR) (Lodh
et al.,
2014).
Constitution of the Primers
Forward and reverse primers were constituted according to the manufacturer’s instructions. Meanwhile,
100 μM of the forward primer was dissolved in 696.52 μl of buffer, while 100 μM of the reverse primer
solution was dissolved in 767.9 μl of buffer solution, the constituted primers were kept at 4°C for further
study (Hany
et al.,
2015)
PCRAmplication of
S. haematobium
PCR was conducted to amplify
S. haematobium
at 121 base pair (bp), the PCR of each sample was
carried out in a 0.2 ml microfuge tube containing 25 μl reaction mix with the following components; 12.5
μl of Top Taq master mix (Qiagen, USA), 6.5 μl of molecular grade water (Qiagen, USA), 0.5 μl of each 10
μM SH-F and SH-R primers and 5 μl of DNA template. The tubes were transferred to Applied Biosystem
(ABS) 9700 Thermocycler and the cycling conditions used were; an initial denaturation at 95°C for 10
mins, followed by 33 cycles of nal denaturation at 94°C for 30 s, annealing at 53°C for 90 s, and
extension at 68°C for 90 s, followed by a nal extension at 60°C for 5 min and post hold at 8°C. The PCR
amplicons were visualized at 1.5% agarose gel (Lodh
et al.,
2014). A list of primers used in the PCR
amplication is provided in Table 3.1.
Table 1: Primers Used for Amplication of
S. haematobium
SN Primers Name Oligonucleotide Sequence
1 SH-F 5’- GATCTCACCTATCAGACGAAAC-3’
2 SH-R 5’-TCACAACGATACGACCAAC-3’
Agarose GelElectrophoresis
Agarose powder of 1g was placed in a conical ask containing 50 ml of buffer, the mixture was
homogenized and microwaved at a medium heat temperature, and the solution was allowed to cool to
room temperature, 3μl of ethidium bromide was placed to pre-stained the gel, the solution was
transferred into the casting plate with an inserted comb and then allowed to solidify within 30 minutes,
the comb was removed from the casting plate, then the casting plate was placed inside the casting
chamber containing 300 ml of buffer solution, the amplied products were subjected to agarose gel
electrophoresis with 100 bp ladder used as standard. The electrophoresis was carried out at 50 volts for
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40 minutes using a BioRad agarose gel electrophoresis unit. The gel was visualized using a U-V trans-
illuminator in a BioRad XRS gel documentation device (Hamdan and Righetti, 2005).
Sodium Dodecyl Sulphate - Polyacrylamide Gel Electrophorese (SDS-PAGE)
Extraction of
S. haematobium
Antigens
Ten millilitres (10 ml) of each positive sample was placed in a test tube and centrifuged at 2000 rpm for
5 minutes, the supernatant was discarded, sediment was suspended in Phosphate Buffer Saline (PBS),
then homogenized and incubated at 4°C for 15 minutes, a small portion of the sample was examined
microscopically to determine the percentage of the disrupted eggs when approximately 95% of the eggs
were disrupted, the crude mixture was centrifuged at 4°C for 20 minutes at 2000 rpm, the supernatant
was collected and ultracentrifuged at 4°C for 90 minutes at 100,000 rpm and then sterilized by passing it
through a 0.2 µm lter, two drops of 2-mercaptoethanol buffer was added into the sample to completely
denature the proteins, each sample was boiled at 95oC for ve (5) minutes then kept into the test tube
for the analysis (Bosompem
et al.,
1996).
Separating Gel Preparation
To ensure proper preparation of 12% Separating gel, 4.35 ml of distilled water, 2.5 ml of Tris buffer, 3.0 ml
of 40% bis-acrylamide solution, 100 microliters of 10% SDS solution, 50 microliters of 10% APS solution,
and 10 microliters of TEMED were measured accordingly and transferred into the same test tube, the
content was homogenized to ensure proper mixture of the resolving gel and stored for further study
(Christine
et al.,
2006).
Stacking Gel Preparation
Prepared 12% stacking gel constituted; 2.95 ml of distilled water, 1.25 ml of tris buffer, 759 µl of 40% bis-
acrylamide solution, 50 microliters of 10% SDS, 30 µl of 10% APS and 8 microliters of TEMED was
measured accordingly and transferred into the same test, the content inside the test tube was
homogenized vigorously to ensure proper mixture of the stacking gel and stored for further study
(Halligan
et al.,
2004).
Gel Electrophoresis Running
Two casting plates was combine together, 1 millimeter thick spacers was use to separate the two
casting plates between the vertical and dawn sizes of the plates, plastic clips were used to tighten the
position of the plates, distilled water was added in to the casting gel to observe linkage, the distilled
water was removed from the casting gel, 12% prepared separating gel solution was poured into the
casting gel until it reached 1 cm away from the top of the short plate, the gel was monitored to ensure
there was no leakages at the top of the plates, 200 microliters of isopropanol was added into the
resolving gel, thin layer was formed on the top of the gel, the separating gel polymerized within 30
minutes, the isopropanol was drained from the top of separating gel, the top of the gel was rinsed gently
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with distilled water and allowed to dry using Kim wipes, the prepared stacking gel solution was poured
on the top of the separating gel, the plastic comb was inserted into the solution between the casting
plates, the gel was allowed to polymerized, the comb was removed from the gel accordingly, distilled
water was used to rinse the holes formed by the comb, the dawn spacer was removed gently from the
casting frame, the gel was inserted vertically into the electrophoresis chamber, tightened accordingly
using a tighten notes inside the electrophoresis chamber, buffer solution was added into the upper and
lower electrophoresis tank chambers respectively, molecular weight ladder was added into the rst hole
and 15 µl of each urine sample was loaded into the subsequent holes accordingly, the anode and
cathode electrodes was set up, electric eld was applied to the gel at 125 volts for 45 minutes,
electrodes were removed, the gel was untighten and removed outside the electrophoresis chamber, the
vertical spacers were removed carefully, the plates were removed from the gel accordingly using gel
remover and subjected for staining (Hamdan and Righetti, 2005).
Gel Staining
Coomassie staining solution was introduced into the staining container, the gel was inserted into the
staining solution and allowed overnight, distilled water was used to rinse the gel and nally molecular
weight of the proteins was observed using a gel documentation device (Kanamura
et al.,
2019).
Western Blotting (WB) Analysis
WB analysis was performed according to the manufacturer’s instructions provided by LD-BIO in a kit
containing all reagents needed for the WB analysis; the protein discovered in the gel were transferred
onto the nitrocellulose sheet, which was coated with Tris-NaCl (pH 7.4), containing 5% nonfat milk, the
blots were washed twice with Tris-NaCl, then dried and incubated with sera diluted in 1:50 Tris-NaCl
sample buffer for 90 mins., after then washed with Tris-NaCl wash buffer, and then incubated with an
anti-human immunoglobulin G alkaline phosphatase conjugate for 60 mins, then washed again, the
proteins fractions recognized by the sera were revealed by the corresponding substrate- a chromogenic
solution containing nitroblue tetrazolium and 5-bromo-4-chloro-3-indolylphosphate. The reaction was
stopped by washing the strips with distilled water and nally, it was dried and placed on a paper for
reading and interpretation (Towbin
et al.,
2019).
Data Analysis
Data obtained was subjected to descriptive and inferential statistics, using International Business
Machine Corporation-Statistical Package for Social Sciences (IBM-SPSS) software tool version 27.0; The
presence of
S. haematobium
and antigen (Sh 22.6 kDa) were considered as dependent variables while
location, age, gender and clinical manifestations were independent variables. Chi-square was used to
identify the association of the infection with location, age, gender and clinical manifestation at P<0.05.
RESULTS
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Prevalence of Sh 22.6 kDa Base on the Location, Gender
and Age of the Participants
Plate 1 showed the bands of Sh 22.6 kDa antigen and Table1 shows the results obtained about
locations, age groups and gender of the participants. Out of 276 infected people examined for the
presence of the Sh 22.6 kDa, 6.2% (17/276) harboured the antigen. Infection with the Sh 22.6 kDa was
not signicantly associated (P = 0.193) with the location. However, the Shagari Centre showed the
highest percentage of the Sh 22.6 antigens [7.1% (6/84)]. Gender is signicantly associated (P = 0.000)
with the antigen infection. Males highly harboured the antigen [14/197 (7.1%)] than females [3.8% (3/79).
The age group is not signicantly associated (P = 0.182) with the infection. However, infected people
aged ≥ 56 years old had the highest Sh 22.6 kDa antigen [40.0% (2/5)] than other age groups.
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Table 2
Prevalence of Sh 22.6 kDa Antigen Among the Infected Individuals
Variables No. Examined No. Positive Prevalence (%) X2 –Value DF P-Value
Locations
Goronyo 43 1 2.3 12.37 3 0.193
Kware 42 3 7.1
Shagari 84 6 7.1
Wamakko 107 7 6.5
Total 276 17 6.2
Gender
Females 79 3 3.8 14.39 1 0.000**
Males 197 14 7.1
Total 276 17 6.2
Age
5 to 15 118 9 7.6 67.59 5 0.184
16 to 25 116 2 1.7
26 to 35 25 2 8.0
36 to 45 8 1 12.5
46 to 55 4 1 25.0
≥ 56 5 2 40.0
Total 276 17 6.2
**= Signicantly Associated DF = Degree of freedom (n-1)
Frequency of Sh 22.6 kDa Antigen with the Clinical
Manifestations
As presented in Table2; Clinical manifestations are associated with Sh 22.6 kDa frequency. Those with
dicult urination experience had the highest signicantly associated (P = 0.000) percentage of the
antigen [(12.6% (16/127)]. Also, those with painful urination had the highest [6.8% (15/221)] frequency.
However, no signicant association (P = 0.064)). Those with experience of bloody urine accounted for
the highest frequency [11.5% (15/131)] with signicant association (P = 0.017). Infected People with
blood-brown urine colour had the highest percentage of the antigen [13/68 (19.1%)], with signicant
association (P = 0.000).
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Table 2
Frequency of the Sh 22.6 Antigen Among the Infected Individuals for the Clinical Manifestations
Clinical Manifestations No.
Examined No.
Positive Prevalence
(%) X2-
Value
DF P-
Value
Dicult Urination 154 1 0.000*
Yes 127 16 12.6
No 149 1 0.7
Total 276 17 6.2
Painful Urine
No 55 2 3.6 2.843 1 0.064
Yes 221 15 6.8
Total 276 17 6.2
Bloody Urine
Experience
Yes 131 15 11.5 36.763 1 0.017*
No 145 2 1.4
Total 276 17 6.2
Physical Observation
Blood Brown 68 13 19.1 25.583 3 0.000*
Dark Brown 22 3 13.6
Light Brown 29 1 3.5
Light Yellow 157 0 0.0
Total 276 17 6.2
Com_B 11 Strip
Haematuria 105 11 10.5 43.761 2 0.083
Proteinuria 97 6 6.2
Normal 74 0 0.0
Total 276 17 6.2
*= Signicantly Associated; DF = Degree of Freedom (n-1)
DISCUSSION
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The present study on the detection of
S. haematobium
antigen (Sh 22.6 kDa) showed that 6.2% of the
infected individuals in the study area, were harbouring
S. haematobium
with the Sh 22.2 kDa. This report
showed a lower prevalence of the antigen among the people in the study area. The low prevalence could
be attributed to the slight behavioural and physiological changes of the infected person in the study area.
Roger et al., (2017) made a similar observation while conducting an expression on analysis of highly
polymorphic mucin proteins (Sm PoMuc) from the parasite of
Schistosoma species;
they reported that
behavioural changes of the infected people may result in the production of resistant antigens from the
parasites due to changes in the IgE response which occurs inside the infected host. The prevalence of
the antigen was lower than the 26.0% identied in studying increased ShTAL1 (sh 22.6kDa) against IgE
before praziquantel treatment in Ghanaians’
S. haematobium
endemic community (Asuming-Brempong
et al., 2022). Also, it was lower than the 35.5% identied by Fitzsimmons et al. (2004) while investigating
human IgE response to the
S. haematobium
22·6 kDa antigen. However, it was in the same range with
10.3% identied in Gabon by Webster
et al.
(2016) in their research on human immunoglobulin E
responses to a recombinant 22.6-kilodalton antigen from
Schistosoma
adult worms are associated with
low intensities of reinfection after treatment.
Age group is signicantly associated with Sh 22.6 kDa frequency in the study area. The highest
frequency of the antigen among the infected people in old age could be due to the capacity of the old
age group (≥ 56 years old) to produce a high amount of IgE which responds to a 22.6-kilodalton antigen
from
Schistosoma
adult worms. The present nding was consistent with earlier research in communities
with endemic
S. mansoni
and
S. haematobium
infections by Dunne et al. (2017), in their research on
immunity after treatment of human schistosomiasis: association between IgE antibodies to adult worm
antigens and resistance to reinfection. Another research by Hagan et al. (2015), on human IgE, IgG4 and
resistance to reinfection with
S. haematobium
also showed that higher production of the resistance
antigen among the
S. haematobium
infected is associated with older age. The current nding did not line
up with earlier research by Dunne et al. (2021), on the isolation of a 22 kDa band after SDS-PAGE of
Schistosoma
adult worms and use to demonstrate that IgE responses against the antigen(s) it contains
are associated with human resistance to reinfection. The researchers said that people with
Schistosoma
infections whose IgE antibodies recognized antigen(s) in the 22-kDa range were less infected with the
antigen.
Gender was signicantly associated with Sh 22.6 kDa distribution among the participants in the present
study. Infected males hosted more antigens than their female counterparts in the study area. This could
be due to frequent males’ exposure to the parasites, which may lead to increased antigen production. A
similar nding was made in an investigation of people who had
S. japonicum
infection in the Philippines
(Webster et al., 1997). Similar to prior results in the other human schistosomiases, it has been
demonstrated that increased IgE titers against worm antigens are favourably linked with gender (Olds
et
al.
, 1997), and the development of gender-specic immunity is supported by the ndings of the current
investigation.
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Clinical manifestations such as bloody urine experience and blood brown urine are signicantly
associated with the prevalence of Sh 22.6 kDa antigen in the study area. This could be attributed to the
fact that infection with
S. haematobium
affects the seminal vesicles, lower ureters, and bladder, the
oxygen gradient created by the urine uid propels the parasites towards the bladder wall, and the
terminal spikes on their eggs are aimed at the bladder lumen when they are laid there, the pathological
lesions, haematuria, proteinuria, dysuria and anuria are caused by the parasite and eggs that are unable
to break free and remain caught in the bladder wall. These eggs release their antigens and cause the
development of granulomas as reported by Dvora et al. (2018). Dunne et al. (2017), reported a similar
observation in their report on immunity after treatment of human schistosomiasis: association between
IgE antibodies to adult worm antigens and resistance to reinfection. Also, Santiago et al. (2018) made
similar observations.
Conclusion
The present study showed that the antigen (Sh 22.6 kDa) started emerging among the
S. haematobium-
infected people in the study area, however, currently, a small proportion of the people (6.2%) harboured
the antigen. The frequency of the antigen was signicantly associated among the participants according
to their gender, age and clinical manifestations and not associated with the location of the participants.
The present survey could serve as baseline data for implementing effective control and preventive
measures in the study area.
Abbreviations
Sh 22.6 kDa =
Schistosoma haematobium
22.6 kilodalton antigen
SDS-PAGE = Sodium Dodecyl Sulphate- Polyacrylamide Gel Electrophoreses
WB= Western Blotting
DNA=Deoxy Ribonucleic Acid
IgE= Immunoglobulin E
DF= Degree of Freedom
X2= Chi-Square
Declarations
Ethics Approval and Consent to Participants
The current study considered all research ethics by the research committee of the Sokoto State Ministry
of Health. The committee approved the research with the letter containing the reference number
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(SKHREC/10/2023). When seeking consent from each individual, the principles, aim, objectives
procedure, and signicance of the study were explained to the participants using the local language
(Hausa), andindividual(s) who were unwilling and those who either refused to return samples were not
included in the survey.
Consent for Publication
Not applicable
Availability of Data Set and Material
Not applicable
Competing Interests
The authors declared no competing interest
Funding
No funds have been received for this research
Authors Contributions
SJ developed the idea of conducting the research, made the initial draft of the research design, searched
for the related literature, participated in sample collection and conducted laboratory work, NTI
supervised the laboratory work and analysed the raw data obtained from the study, EVU designed the
questionnaire, sort for validity and reliability test of the questionnaire; BIA sort for the ethical approval,
partake in sample collection and supervision of the laboratory analysis work.
Acknowledgements
The authors acknowledged: Professor Fahrul Zaman Huyop of the Biosciences Department, Faculty of
Science, Universiti Teknologi Malaysia; Sokoto State Ministry of Health; District Heads and Riverine
Community members for their countless contributions to this research to be carried out.
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Figures
Figure 1
