Emergence of Sh 22.6 kDa Antigen Among the Schistosoma haematobium Infected People in Riverine Communities of the Extreme North Western Nigeria

Emergence of Sh 22.6 kDa Antigen Among the Schistosoma haematobium Infected People in Riverine Communities of the Extreme North Western Nigeria | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Emergence of Sh 22.6 kDa Antigen Among the Schistosoma haematobium Infected People in Riverine Communities of the Extreme North Western Nigeria Suleman Jafaru, Isyaku Nike Tawakaltu, Ukatu Victoria Ebere, Bagudo Ahmad Ibrahim This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4843645/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 5 You are reading this latest preprint version 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 filtration method, all microscopy-positive samples were subjected to PCR analysis for confirmation 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 significantly 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 significant association (P = 0.000). Although, the age group is not significantly 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 fight against urinary schistosomiasis in the area. Antigen Emergence Filtration Technique PCR Riverine Communities Schistosoma haematobium SDS-PAGE Urine Figures Figure 1 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 deficiency 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 ). 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 identification 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 efficacy 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 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 floodplains of the Sokoto-Rima River system, the state is also covered with rich alluvial 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 five (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 identified 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 significance, 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 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 field 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), difficult 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 4 0 C and transported to the parasitology laboratory, Department of Zoology, Usmanu Danfodiyo University, Sokoto for the analysis (Afifi et al., 2016). Filtration of the Collected Urine Samples and Microscopy Each sample was filtered using the standard method recommended by WHO (2002). The sample was filtered through Whatman filter paper 0.5mm using a filtration machine within five minutes, each filter 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 filter 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% 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 finally, 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) PCR Amplification 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 final 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 final 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 amplification is provided in Table 3.1. Table 1: Primers Used for Amplification of S. haematobium SN Primers Name Oligonucleotide Sequence 1 SH-F 5’- GATCTCACCTATCAGACGAAAC-3’ 2 SH-R 5’-TCACAACGATACGACCAAC-3’ Agarose Gel Electrophoresis Agarose powder of 1g was placed in a conical flask 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, 3 μl 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 amplified products were subjected to agarose gel electrophoresis with 100 bp ladder used as standard. The electrophoresis was carried out at 50 volts for 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 filter, two drops of 2-mercaptoethanol buffer was added into the sample to completely denature the proteins, each sample was boiled at 95 o C for five (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 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 first hole and 15 µl of each urine sample was loaded into the subsequent holes accordingly, the anode and cathode electrodes was set up, electric field 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 finally 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 finally, 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 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 Table 1 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 significantly 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 significantly 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 significantly 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. Table 2 Prevalence of Sh 22.6 kDa Antigen Among the Infected Individuals Variables No. Examined No. Positive Prevalence (%) X 2 –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 **= Significantly Associated DF = Degree of freedom (n-1) Frequency of Sh 22.6 kDa Antigen with the Clinical Manifestations As presented in Table 2 ; Clinical manifestations are associated with Sh 22.6 kDa frequency. Those with difficult urination experience had the highest significantly 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 significant association (P = 0.064)). Those with experience of bloody urine accounted for the highest frequency [11.5% (15/131)] with significant association (P = 0.017). Infected People with blood-brown urine colour had the highest percentage of the antigen [13/68 (19.1%)], with significant association (P = 0.000). Table 2 Frequency of the Sh 22.6 Antigen Among the Infected Individuals for the Clinical Manifestations Clinical Manifestations No. Examined No. Positive Prevalence (%) X 2 -Value DF P-Value Difficult 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 *= Significantly Associated; DF = Degree of Freedom (n-1) DISCUSSION 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% identified 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% identified 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% identified 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 significantly 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 finding 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 finding 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 significantly 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 finding 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-specific immunity is supported by the findings of the current investigation. Clinical manifestations such as bloody urine experience and blood brown urine are significantly 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 fluid 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 Dvorǎ 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 significantly 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 X 2 = 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 (SKHREC/10/2023). When seeking consent from each individual, the principles, aim, objectives procedure, and significance of the study were explained to the participants using the local language (Hausa), and individual(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. References Afifi, M., Jiman-Fatani, A., Al-Hussainy, N., Al-Rabia, M. & Bogari, A. (2016). Genetic diversity among natural populations of Schistosoma haematobium might contribute to inconsistent virulence and diverse clinical outcomes. Journal of Microscopic Ultrastructures , 4 (4): 222-229. https://doi.org/10.1038/s41598-022-11645-3 Agi, P. I., & Okafor, E. J. (2005). The epidemiology of Schistosoma haematobium in Odau community in the Niger Delta Area of Nigeria. Journal of Applied Science and Environmental Management . 9 (3): 37-43. Arshad, M. & Odeh, G. O. (2014). The Establishment of the Nigerian Sokoto Caliphate: An inquest into the Background History of the 1804 Jihad in Hausa Land, 210 years After, Al-Qalam Journal of Arts and Social Sciences, 2 (1): 61-79 Asmaa, M. El-Kady, M., EL-Amir, M. H. H., Khaled, S. A., Ahmad, A. & Alzahraa, A. A. (2020). Genetic Diversity of Schistosoma haematobium in Qena Governorate, Upper Egypt. Infection and Drug Resistance, 13 (2): 3601–3611 Asuming-Brempong, E. K., Ayi, I., van der Puije, W., Gyan, B. A., Larbi, I. A., Ashong, Y., Frempong, N. A., Quartey, J. K., Otchere, J., Jones, F. M., Wilson, S., Dunne, D. W., & Boakye, D. A. (2022). Increased ShTAL1 IgE responses post-Praziquantel treatment may be associated with a reduced risk of re-infection in a Ghanaian S. haematobium-endemic community. PLoS Neglected Tropical Diseases , 16 (3). https://doi.org/10.1371/journal.pntd.0010115 Atalabi, T. E., Lawal, U., & Ipinlaye, S. J. (2016). Prevalence and intensity of genito-urinary schistosomiasis and associated risk factors among junior high school students in two local government areas around Zobe Dam in Katsina State, Nigeria. Parasites and Vectors , 9 (1). https://doi.org/10.1186/s13071-016-1672-5 Black, C. L., Mwinzi, P. N. M., Muok, E. M. O., Abudho, B., Fitzsimmons, C. M., Dunne, D. W., Karanja, D. M. S., Secor, W. E., & Colley, D. G. (2010). Influence of exposure history on the immunology and development of resistance to human Schistosomiasis mansoni. PLoS Neglected Tropical Diseases , 4 (3). https://doi.org/10.1371/journal.pntd.0000637 Bosompem, K. M., Arishima, T., Yamashita, T., Ayi, I., Anyan, W. K., & Kojima, S. (1996). Extraction of Schistosoma haematobium antigens from infected human urine and generation of potential diagnostic monoclonal antibodies to urinary antigens . Acta Tropica, 62 (2), 91–103 . doi:10.1016/s0001-706x(96)00040-x Casacuberta-Partal, M., Janse, J. J., van Schuijlenburg, R., de Vries, J. J. C., Erkens, M. A. A., Suijk, K., van Aalst, M., Maas, J. J., Grobusch, M. P., van Genderen, P. J. J., de Dood, C., Corstjens, P. L. A. M., van Dam, G. J., van Lieshout, L., & Roestenberg, M. (2021). Antigen-based diagnosis of Schistosoma infection in travellers: A prospective study. Journal of Travel Medicine , 27 (4). https://doi.org/10.1093/JTM/TAAA055 Christine, P., Shona, W., He´le`ne, L., Sophia, L., Frances, M. J., Wassim, D., Monique, C., David, W. D. & Jamal, K. (2006). Identification of a Novel Antigen of Schistosoma mansoni Shared with Plasmodium falciparum and Evaluation of Different Cross-Reactive Antibody Subclasses Induced by Human Schistosomiasis and Malaria. Infection and Immunity , 74 (6) 3347–3354. Daumerie, Denis., Savioli, Lorenzo., Crompton, D. W. T. (David W. T., & World Health Organization. Department of Control of Neglected Tropical Diseases. (2010). Working to overcome the global impact of neglected tropical diseases: first WHO report on neglected tropical diseases. World Health Organization Di Bella, S., Riccardi, N., Giacobbe, D. R., & Luzzati, R. (2018). History of schistosomiasis (bilharziasis) in humans: from Egyptian medical papyri to molecular biology on mummies. Pathogens and Global Health , 112 (5), 268–273. https://doi.org/10.1080/20477724.2018.1495357 Dunne, D. W., Butterworth, A. E., & Fulford, A. J. (2017). Immunity after treatment of human schistosomiasis: association between IgE antibodies to adult worm antigens and resistance to reinfection. European Journal Immunology. 22 : 1483–1494. Dunne, D. W., Webster, M., Smith, P., Langley, J. G., Richardson, B. A., Fulford, A. J. C., Butterworth, A. E., Sturrock, R. F., Kariuki, H. C. & Ouma, J. H. (2021): The isolation of a 22 kDa band after SDS-PAGE of Schistosoma mansoni adult worms and its use to demonstrate that IgE responses against the antigen(s) it contains are associated with human resistance to reinfection. Parasite Immunology. 19:79–89. Dvorǎ, ́K. J., Mashiyama, S. T., Braschi, S., Sajid, M., Knudsen, G. M., & Hansell, E, (2018). Differential use of protease families for invasion by schistosome cercariae. Biochemistry. 90 (2):345–358 El-Kady, A. M., El-Amir, M. I., Hassan, M. H., Allemailem, K. S., Almatroudi, A., & Ahmad, A. A. (2020). Genetic diversity of Schistosoma haematobium in Qena governorate, Upper Egypt. Infection and Drug Resistance , 13 , 3601–3611. https://doi.org/10.2147/IDR.S266928 El-Kady, A. M., El-Amir, M. I., Hassan, M. H., Allemailem, K. S., Almatroudi, A., & Ahmad, A. A. (2020). Genetic diversity of Schistosoma haematobium in Qena governorate, Upper Egypt. Infection and Drug Resistance , 13 , 3601–3611. https://doi.org/10.2147/IDR.S266928 Fitzsimmons, C. M., Stewart, T. J., Hoffmann, K. F., Grogan, J. L., Yazdanbakhsh, M., & Dunne, D. W. (2004). Human IgE response to the Schistosoma haematobium 22·6 kDa antigen. In Parasite Immunology (Vol. 26). www.sanger.ac.uk Galazka, A. M. (1993). The Immunological Basis for Immunization Series General Immunology Medical Officer Expanded Programme on Immunization global programme for vaccines and immunization expanded programme on immunization The Expanded Programme on Immunization thanks the following donors whose support made the production of these modules possible: United Nations Development Fund (UNDP) The Rockefeller Foundation . http://www.who.ch/programmes/gpv/gEnglish/avail/gpvcatalog/catlog1.htm Gamde, S. M., Garba, A., Tongvwam, P., Gamde, S. D., Agom, D. D., & Igbinosa, I. (2023). Prevalence, intensity, and risk factors of urinary schistosomiasis among primary school children in Silame, Sokoto, Nigeria. Microbes and Infectious Diseases , 4 (2), 695–703. https://doi.org/10.21608/MID.2022.155035.1365 Hagan P, Blumenthal UJ, Dunne D, Simpson AJG, Wilkins HA (2015): Human IgE, IgG4 and resistance to reinfection with Schistosoma haematobium . Nature ; 349 :243–245. Halligan, B.D., Rootti, V., Jin, W., Laffoo, S., Twigger, S.N., & Dratz, E.A., (2004). ProMoSt [protein Modification screening tool] is a web-based tool for mapping protein modification of two-dimensional gels. Nucleic Acids Research, 32: 638-644. Hamdan, M., & Righetti, P. G., (2005). John Wiley & Sons, Inc., Proteomics Today: Protein Assessment and Biomarkers Using Mass Spectrometry, Electrophoresis, and Microarray Technology, Hoboken, Nature Journal, USA. 274–340. Hany Sady, Hesham M. Al-Mekhlafi, Romano Ngui, Wahib M. Atroosh, Ahmed K. Al-Delaimy, Nabil A. Nasr, Salwa Dawaki, Awatif M. Abdulsalam, Init Ithoi, Yvonne A. L. Lim, Kek Heng Chua, & Johari, S. (2015). Detection of Schistosoma mansoni and Schistosoma haematobium by PCR with High-Resolution Melting Analysis International Journal of Molecular Science. 16 , 16085-16103 Ismail, A., & Oke, I. (2012). Statistical and trend analyses of rainfall in Sokoto. International Research Journal of Engineering Science, Technology and Innovation , 1 : 161–174 Jibrillah, A. M., Ja’afar, M., & Choy, L. K. (2019). Monitoring Vegetation Change in the Dryland Ecosystem of Sokoto, Northwestern Nigeria using Geoinformatics. Indonesian Journal of Geography , 51 (1): 9–17. Jiya, F. B., Jiya, N. M., Ibitoye, P. K., Mohammed, Y., & Mohammad, A. U. (2022). Chronic Urogenital Schistosomiasis among In-School Adolescents in Sokoto, North-Western Nigeria. Asian Journal of Medicine and Health , 22–36. https://doi.org/10.9734/ajmah/2022/v20i130429 Kanamura, H. Y., Hoshino-Shimizu, S., Camargo, M. E. & Silva, L. C. (2019). Class-specific antibodies and fluorescent staining patterns in acute and chronic forms of Schistosoma mansoni. American Journal and Tropical Medicine and Hygiene ; 28: 242-248 Kasim, I. & Usman, A. Y. (2021). Trend analysis of Temperature and Precipitation Change in Sokoto State, Nigeria Journal of Agricultural Economics and Rural Development. 7 (1): 941-951 Ladan, M. U., Abubakar, U., Abdullahi, K., Bunza, M. D. A., Ladan, M. J., & Adamu, T. (2012). Studies on Urinary Schistosomiasis in Selected Villages around Gusau Dam Site, Zamfara State, Nigeria. Nigerian Journal of Basic and Applied Science , 20 (3), 189–194. http://www.ajol.info/index.php/njbas/index Lodh N, Naples, J. M., Bosompem, K. M., Quartey, J. & Shiff, C. J. (2014). Detection of Parasite-Specific DNA in Urine Sediment Obtained by Filtration Differentiates between Single and Mixed Infections of Schistosoma mansoni and S. haematobium from Endemic Areas in Ghana. PLoS ONE. 9 (3): e91144. Markakpo, U. S., Armah, G. E., Fobil, J. N., Asmah, R. H., Anim-Baidoo, I., Dodoo, A. K., Madjitey, P., Essuman, E. E., Kojima, S., & Bosompem, K. M. (2015). Immunolocalization of the 29 kDa Schistosoma haematobium species-specific antigen: A potential diagnostic marker for urinary schistosomiasis. BMC Infectious Diseases , 15 (1). https://doi.org/10.1186/s12879-015-0931-y Mohammed, I. H., Mohammed, H. I., Yahaya, I., & Buhari, S. B. (2020). Prevalence and Determining Factors of Urinary Schistosomiasis among Primary School Pupils in North Central Nigeria. In Asian Journal of Parasitology (Vol. 4, Issue 3). Mwai, J., Njenga, S., & Barasa, M. (2016). Knowledge, attitude and practices concerning prevention and control of schistosomiasis infection in Mwea Kirinyaga county, Kenya. In BMC Public Health (Vol. 16, Issue 1). BioMed Central Ltd. https://doi.org/10.1186/s12889-016-3494-y Olds GR, Olveda R, Wu G, Wiest P, McGarvey S, Aligui G, Zhang S, Ramirez B, Daniel B, Peters P, Romulo R, Fevidal P, Tiu W, Yuan J, Domingo E. & Blas B (1996): Immunity and morbidity in schistosomiasis japonicum infection. American Journal of Tropical Medicine Hygiene ; 55 (5):121–126. Pearson, M. S., Tedla, B. A., Mekonnen, G. G., Proietti, C., Becker, L., Nakajima, R., Jasinskas, A., Doolan, D. L., Amoah, A. S., Knopp, S., Rollinson, D., Ali, S. M., Kabole, F., Hokke, C. H., Adegnika, A. A., Field, M. A., van Dam, G., Corstjens, P. L. A. M., Mduluza, T. & Loukas, A. (2021). Immunomics-guided discovery of serum and urine antibodies for diagnosing urogenital schistosomiasis: a biomarker identification study. The Lancet Microbe , 2 (11), e617–e626. https://doi.org/10.1016/S2666-5247(21)00150-6 Pinto-Almeida, A., Mendes, T., de Oliveira, R. N., Corrêa, S. de A. P., Allegretti, S. M., Belo, S., Tomás, A., Anibal, F. de F., Carrilho, E., & Afonso, A. (2016). Morphological characteristics of Schistosoma mansoni PZQ-resistant and -susceptible strains are different in the presence of Praziquantel. Frontiers in Microbiology , 7 (APR). https://doi.org/10.3389/fmicb.2016.00594 Roger E, Gourbal B, Grunau C, Pierce RJ, Galinier R, & Mitta G. (2017). Expression analysis of highly polymorphic mucin proteins (Sm PoMuc) from the parasite Schistosoma mansoni. Molecular Biochemistry and Parasitology. 157 (2):217– 27. Santiago, M. L. Hafalla, J. C. & Kurtis J. D. (2018)., “Identification of the Schistosoma japonicum 22.6-kDa antigen as a major target of the human IgE response: similarity of IgE-binding epitopes to allergen peptides,” International Archives of Allergy and Immunology , 117( 2): 94–104. Santos, L. L., Santos, J., Gouveia, M. J., Bernardo, C., Lopes, C., Rinaldi, G., Brindley, P. J., & da Costa, J. M. C. (2021). Review urogenital schistosomiasis—history, pathogenesis, and bladder cancer. In Journal of Clinical Medicine (Vol. 10, Issue 2, pp. 1–11). MDPI. https://doi.org/10.3390/jcm10020205 Senghor, B., Diaw, O. T., Doucoure, S., Sylla, S. N., Seye, M., Talla, I., Bâ, C. T., Diallo, A., & Sokhna, C. (2015). Efficacy of praziquantel against urinary schistosomiasis and reinfection in Senegalese school children where there is a single well-defined transmission period. Parasites and Vectors , 8 (1). https://doi.org/10.1186/s13071-015-0980-5 Sepulveda, J., Tremblay, J. M., DeGnore, J. P., Skelly, P. J., & Shoemaker, C. B. (2010). Schistosoma mansoni host-exposed surface antigens characterized by sera and recombinant antibodies from schistosomiasis-resistant rats. International Journal for Parasitology , 40 (12), 1407–1417. https://doi.org/10.1016/j.ijpara.2010.04.019 Singh, K., Muddasiru, D., & Singh, J. (2016). Current status of schistosomiasis in Sokoto, Nigeria. Parasite Epidemiology and Control , 1 (3), 239–244. https://doi.org/10.1016/j.parepi.2016.08.003 Suleiman, J., T. Isyaku, N., E. Ukatu, V., & Alhaji, Y. I. (2022). Antigens of Three Medically Important Schistosoma spp. Asian Journal of Research in Zoology , 11–27. https://doi.org/10.9734/ajriz/2022/v5i330139 Towbin, H., Staehlin, T. and Gordon, J. (2019). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Procedures for National Academic Science, 76 : 4350–4354. Vale, N., Gouveia, M. J., Rinaldi, G., Brindley, P. J., Gärtner, F., & Da Costa, J. M. C. (2017). Praziquantel for schistosomiasis: Single-drug metabolism revisited, mode of action, and resistance. In Antimicrobial Agents and Chemotherapy (Vol. 61, Issue 5). American Society for Microbiology. https://doi.org/10.1128/AAC.02582-16 Vinkeles, N., Melchers, V. S., van Dam, G. J., Shaproski, D., Kahama, A. I., Brienen, E. A. T., Vennervald, B. J., & van Lieshout, L. (2014). Diagnostic Performance of Schistosoma Real-Time PCR in Urine Samples from Kenyan Children Infected with Schistosoma haematobium : Day-to-day Variation and Follow-up after Praziquantel Treatment. PLoS Neglected Tropical Diseases , 8 (4). https://doi.org/10.1371/journal.pntd.0002807 Webster, M., Ramirez, B. L., Aligui, G. D., Olveda, R. M., Ouma, J. H., Kariuki, H. C., Kimani, G., Olds, G. R., Fulford, J. C., Butterworth, A. E. & Dunne, D. W. (1997). The influence of sex and age on antibody isotype responses to Schistosoma mansoni and Schistosoma japonicum in human populations in Kenya and the Philippines. Parasitology ; 114:383–393 World Health Organization (2002). Prevention and control of schistosomiasis and soil-transmitted helminthiasis. 2003/02/21 ed. Geneva: World Health Organization ; i–vi, 1–57, back cover p Supplementary Files floatimage2.jpeg floatimage3.jpeg Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Minor revisions needed 17 Jul, 2025 Reviewers agreed at journal 09 Jun, 2025 Reviewers invited by journal 05 Aug, 2024 Editor assigned by journal 02 Aug, 2024 First submitted to journal 01 Aug, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4843645","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":336086500,"identity":"f997dfcd-f3a6-4460-a8c5-334cfe8f0856","order_by":0,"name":"Suleman Jafaru","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABHUlEQVRIiWNgGAWjYFAC5gaGBDDjAMMHBgYbiCAPXi2McC2MMxgY0ojUAmMBtRwmrEV+RmLjg4c5dnkMjIcfNvzcc95efkYC44O3bQx58g7YtRjcSGw2SNyWXMzAcMywsefZ7cQNNxKYDee2MRQbHsChRSKxTSJxG3NiA8MB8wc8B24nGEgksEnztjEkbmzArgXosPYfidvqgVqOf2z8c+AcyGHsv/FpYbiRCJTddhio5YxhM8+BA4wNNxLYmEFa5uPQYXDmYTPQYceBGs8UNsscSE7cABSRnHNOInEDrhBrTz748ee26sR+ieMbG98csLMHiXx4U2aTOB+Xw2CATeIAjAmOKQkGgwO41MIAP7qh8oRsGQWjYBSMgpECAIfGZrHJ+pFRAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0001-9486-0797","institution":"Sokoto State University","correspondingAuthor":true,"prefix":"","firstName":"Suleman","middleName":"","lastName":"Jafaru","suffix":""},{"id":336086501,"identity":"317be307-c3fe-4dea-aaf7-534224ba1b5f","order_by":1,"name":"Isyaku Nike Tawakaltu","email":"","orcid":"","institution":"Kebbi State University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Isyaku","middleName":"Nike","lastName":"Tawakaltu","suffix":""},{"id":336086502,"identity":"97887c38-7841-4e3f-b3f8-ff8815cc9b88","order_by":2,"name":"Ukatu Victoria Ebere","email":"","orcid":"","institution":"Kebbi State University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Ukatu","middleName":"Victoria","lastName":"Ebere","suffix":""},{"id":336086503,"identity":"3eaf7b5c-2a31-4258-8ebe-31b635d88542","order_by":3,"name":"Bagudo Ahmad Ibrahim","email":"","orcid":"","institution":"Kebbi State University of Science and Technology","correspondingAuthor":false,"prefix":"","firstName":"Bagudo","middleName":"Ahmad","lastName":"Ibrahim","suffix":""}],"badges":[],"createdAt":"2024-08-01 17:16:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4843645/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4843645/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":63601217,"identity":"effd5c6a-2dc3-4ec4-a57b-d8808039b490","added_by":"auto","created_at":"2024-08-30 05:24:31","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":252855,"visible":true,"origin":"","legend":"\u003cp\u003eMap of Sokoto State Showing the Study Areas (GIS, 2022)\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-4843645/v1/dc89f3900cfd24de51c0b53d.png"},{"id":63601223,"identity":"f60514b5-44fe-4438-b248-caea2b3a23f1","added_by":"auto","created_at":"2024-08-30 05:24:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1149821,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4843645/v1/07988198-8448-4890-9fba-5c223603968d.pdf"},{"id":63601221,"identity":"b3144809-782a-49a2-8c40-57047dc8600a","added_by":"auto","created_at":"2024-08-30 05:24:31","extension":"jpeg","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":170057,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4843645/v1/5368289ef3a9f6ca56d83533.jpeg"},{"id":63601220,"identity":"bd90ac83-d263-4969-85db-97b0e8779780","added_by":"auto","created_at":"2024-08-30 05:24:31","extension":"jpeg","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":110033,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4843645/v1/6ad634d1b06b5e94ebd358ef.jpeg"}],"financialInterests":"","formattedTitle":"Emergence of Sh 22.6 kDa Antigen Among the Schistosoma haematobium Infected People in Riverine Communities of the Extreme North Western Nigeria","fulltext":[{"header":"Background","content":"\u003cp\u003eThe most dangerous species of schistosomes in Nigeria is \u003cem\u003eS. haematobium\u003c/em\u003e, which can lead to severe renal complications affecting cognitive domain and urinary tract pathology (Santos et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Apart from pathology, there is evidence that \u003cem\u003eS. haematobium\u003c/em\u003e can affect nutritional status and lower levels of haemoglobin (Hb) by decreasing food intake and increasing blood loss, which can result in iron deficiency anaemia (Atalabi et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). 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., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), However, the disease is still emerging and reemerging in the nation (Ladan et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Mohammed et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Jiya et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAntigens 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., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Antigens typically consist of a large number of determinants that may repeat molecular structures or be distinct from one another (Sepulveda et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Casacuberta-Partal et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Suleiman et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). 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., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). 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., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Due to its greater capacity to quickly produce various resistant antigens on the surface of its teguments, \u003cem\u003eS. haematobium\u003c/em\u003e induces varying degrees of resistance (El-Kady et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAn 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., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Numerous global investigations examined the existence and roles of 22.6 kDa antigen in \u003cem\u003eSchistosoma species\u003c/em\u003e (Markakpo et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). It was reported that in \u003cem\u003eS. mansoni\u003c/em\u003e, human IgE response against 22\u0026middot;6 kDa antigen known as Sm 22\u0026middot;6 kDa (Fitzsimmons et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Research in Kenya indicated that IgE to Sm 22\u0026middot;6 may contribute to resistance or be a marker of resistant parasites (Galazka, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Mwai et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Pearson et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Another research on \u003cem\u003eS. japonicum\u003c/em\u003e (Sj 22\u0026middot;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., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Also, another on \u003cem\u003eS. haematobium\u003c/em\u003e antigen (Sh 22\u0026middot;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 \u003cem\u003eet al.\u003c/em\u003e, 2005), Kwale district of Kenya (Vinkeles et al., \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), Senegal (Senghor et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) and Egypt (Di Bella et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e); also, the resistance may likely arise when the parasites produce the resistant genes or antigen (Pinto-Almeida et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eS. haematobium\u003c/em\u003e was the most re-emergent species among the \u003cem\u003eSchistosoma species\u003c/em\u003e in the riverine communities of Sokoto (Singh et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Despite its record as the most pathogenic species of schistosomes in Nigeria (El-Kady et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). However, the reports on whether the antigens were among the factors increasing the reoccurrence in the area were scanty. Therefore, the identification 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 efficacy 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.\u003c/p\u003e "},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eStudy Area\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eRiverine 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\u0026deg; 0\u0026apos; 21.1428\u0026apos;\u0026apos; N and 5\u0026deg; 14\u0026apos; 51.1872\u0026apos;\u0026apos; 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\u0026apos;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\u0026deg;C (82.9 \u0026deg;F). Although the area is generally very hot, the dryness helps keep the highest temperatures below 40\u0026deg;C (104.0\u0026deg;F) for most of the year. The warmest months are February to April when daytime temperatures can exceed 45\u0026deg;C (113.0\u0026deg;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 \u003cem\u003eet al.,\u003c/em\u003e 2019).\u003c/p\u003e\n\u003cp\u003eHausa/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).\u0026nbsp;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 floodplains of the Sokoto-Rima River system, the state is also covered with rich\u0026nbsp;alluvial 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).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStudy Population\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study population for the present research consisted of \u003cem\u003eS. haematobium\u003c/em\u003e-infected people aged five (5) years and above residing in the riverine communities of Sokoto State.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResearch Design\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA cross-sectional research design involving a simple random sampling technique was employed in the present research. Meanwhile, out of the identified Local Governments with riverine communities, four (4) Local Governments (Wamakko, Kware, Goronyo, and Shagari) were assigned as sample collection centres based on convenience.\u0026nbsp;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 significance, 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 participant, the samples were subjected to physical observations, and then analyzed in the laboratory (Agi and Okafor, 2005).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eQuestionnaire Administration\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA 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 \u003cem\u003eet al.,\u003c/em\u003e 2020).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Collection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eQualitative and quantitative data were collected from the field and laboratory during the present study. The qualitative data collected were place of living, gender (males and females), presence of \u003cem\u003eS. haematobium\u0026nbsp;\u003c/em\u003e(Yes or No), difficult 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 \u003cem\u003eS. haematobium\u003c/em\u003e eggs per 10ml of urine sample) and age of the participants.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCollection of Urine Sample\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFifty 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\u0026rsquo;s questionnaire. Each collected sample was maintained at 4\u003csup\u003e0\u003c/sup\u003eC and transported to the parasitology laboratory, Department of Zoology, Usmanu Danfodiyo University, Sokoto for the analysis (Afifi \u003cem\u003eet al.,\u003c/em\u003e 2016).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFiltration\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eof the Collected Urine Samples and Microscopy\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEach sample was filtered using the standard method recommended by WHO (2002). The sample was filtered through Whatman filter paper 0.5mm using a filtration machine within five minutes, each filter paper was transferred onto the carbon paper, and three drops of ninhydrin and two drops of Logu\u0026rsquo;s iodine solutions were added respectively on the contents of the filter paper, the paper was allowed to dry at room temperature and observed microscopically using X10 the X40 objective lens.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDNA Extraction\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDeoxyribose 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 \u0026mu;l) was placed into a DNA extraction tube (Microtube), 300 \u0026mu;l of lysis buffer was added into the tube, another 500 \u0026mu;l 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 \u0026nbsp;for the second time, subsequently, 500 \u0026mu;l of wash buffer (70% 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 \u0026mu;l 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 \u0026mu;l of molecular grade water was added and centrifuged at 13,000 rpm for 1 minute, the supernatant was discarded and finally, the extracted DNA was stored for the polymerase chain reaction (PCR) (Lodh \u003cem\u003eet al.,\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e2014).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConstitution of the Primers\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eForward and reverse primers were constituted according to the manufacturer\u0026rsquo;s instructions. Meanwhile, 100 \u0026mu;M of the forward primer was dissolved in 696.52 \u0026mu;l of buffer, while 100 \u0026mu;M of the reverse primer solution was dissolved in 767.9 \u0026mu;l of buffer solution, the constituted primers were kept at 4\u0026deg;C for further study (Hany \u003cem\u003eet al.,\u003c/em\u003e 2015)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePCR\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eAmplification of \u003cem\u003eS. haematobium\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePCR was conducted to amplify \u003cem\u003eS. haematobium\u003c/em\u003e at 121 base pair (bp), the PCR of each sample was carried out in a 0.2 ml microfuge tube containing 25 \u0026mu;l reaction mix with the following components; 12.5 \u0026mu;l of Top Taq master mix (Qiagen, USA), 6.5 \u0026mu;l of molecular grade water (Qiagen, USA), 0.5 \u0026mu;l of each 10 \u0026mu;M SH-F and SH-R primers and 5 \u0026mu;l of DNA template. The tubes were transferred to Applied Biosystem (ABS) 9700 Thermocycler and the cycling conditions used were; an initial denaturation at 95\u0026deg;C for 10 mins, followed by 33 cycles of final denaturation at 94\u0026deg;C for 30 s, annealing at 53\u0026deg;C for 90 s, and extension at 68\u0026deg;C for 90 s, followed by a final extension at 60\u0026deg;C for 5 min and post hold at 8\u0026deg;C. The PCR amplicons were visualized at 1.5% agarose gel (Lodh \u003cem\u003eet al.,\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e2014). A list of primers used in the PCR amplification is provided in Table 3.1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1:\u003c/strong\u003e Primers Used for Amplification of \u003cem\u003eS. haematobium\u003c/em\u003e\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"548\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.02919708029197%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSN\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"26.45985401459854%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePrimers Name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"65.51094890510949%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eOligonucleotide Sequence\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.02919708029197%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"26.45985401459854%\" valign=\"top\"\u003e\n \u003cp\u003eSH-F\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"65.51094890510949%\" valign=\"top\"\u003e\n \u003cp\u003e5\u0026rsquo;- GATCTCACCTATCAGACGAAAC-3\u0026rsquo;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"8.02919708029197%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"26.45985401459854%\" valign=\"top\"\u003e\n \u003cp\u003eSH-R\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"65.51094890510949%\" valign=\"top\"\u003e\n \u003cp\u003e5\u0026rsquo;-TCACAACGATACGACCAAC-3\u0026rsquo;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003eAgarose Gel\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eElectrophoresis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAgarose powder of 1g was placed in a conical flask 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, 3\u0026nbsp;\u0026mu;l 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 amplified products were subjected to agarose gel electrophoresis with 100 bp ladder used as standard. The electrophoresis was carried out at 50 volts for 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).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSodium Dodecyl Sulphate - Polyacrylamide Gel Electrophorese (SDS-PAGE)\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExtraction of \u003cem\u003eS. haematobium\u0026nbsp;\u003c/em\u003eAntigens\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTen 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\u0026deg;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\u0026deg;C for 20 minutes at 2000 rpm, \u0026nbsp;the supernatant was collected and ultracentrifuged at 4\u0026deg;C for 90 minutes at 100,000 rpm and then sterilized by passing it through a 0.2 \u0026micro;m filter, two drops of 2-mercaptoethanol buffer was added into the sample to completely denature the proteins, each sample was boiled at 95\u003csup\u003eo\u003c/sup\u003eC for five (5) minutes then kept into the test tube for the analysis (Bosompem\u003cem\u003e\u0026nbsp;et al.,\u003c/em\u003e1996).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSeparating Gel Preparation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo 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 \u003cem\u003eet al.,\u003c/em\u003e 2006).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStacking Gel Preparation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePrepared 12% stacking gel constituted; 2.95 ml of distilled water, 1.25 ml of tris buffer, 759 \u0026micro;l of 40% bis-acrylamide solution, 50 microliters of 10% SDS, 30 \u0026micro;l 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 \u003cem\u003eet al.,\u003c/em\u003e 2004).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGel Electrophoresis Running\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTwo 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 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 first hole and 15 \u0026micro;l of each urine sample was loaded into the subsequent holes accordingly, the anode and cathode electrodes was set up, electric field 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).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGel Staining\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCoomassie 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 finally molecular weight of the proteins was observed using a gel documentation device (Kanamura \u003cem\u003eet al.,\u003c/em\u003e 2019).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eWestern Blotting (WB) Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWB analysis was performed according to the manufacturer\u0026rsquo;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 finally, it was dried and placed on a paper for reading and interpretation (Towbin \u003cem\u003eet al.,\u003c/em\u003e 2019).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData 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 \u003cem\u003eS. haematobium\u003c/em\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003eand 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\u0026lt;0.05.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003ePrevalence of Sh 22.6 kDa Base on the Location, Gender and Age of the Participants\u003c/h2\u003e \u003cp\u003ePlate 1 showed the bands of Sh 22.6 kDa antigen and Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e 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 significantly associated (P\u0026thinsp;=\u0026thinsp;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 significantly associated (P\u0026thinsp;=\u0026thinsp;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 significantly associated (P\u0026thinsp;=\u0026thinsp;0.182) with the infection. However, infected people aged\u0026thinsp;\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;56 years old had the highest Sh 22.6 kDa antigen [40.0% (2/5)] than other age groups.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrevalence of Sh 22.6 kDa Antigen Among the Infected Individuals\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariables\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. Examined\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNo. Positive\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePrevalence (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eX\u003csup\u003e2\u003c/sup\u003e \u0026ndash;Value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eDF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eP-Value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLocations\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGoronyo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.193\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKware\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eShagari\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWamakko\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e107\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGender\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemales\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.000**\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMales\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e197\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAge\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5 to 15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e118\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e67.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.184\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16 to 25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e116\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e26 to 35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e36 to 45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e46 to 55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e25.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e**= Significantly Associated DF\u0026thinsp;=\u0026thinsp;Degree of freedom (n-1)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eFrequency of Sh 22.6 kDa Antigen with the Clinical Manifestations\u003c/h2\u003e \u003cp\u003eAs presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e2\u003c/span\u003e; Clinical manifestations are associated with Sh 22.6 kDa frequency. Those with difficult urination experience had the highest significantly associated (P\u0026thinsp;=\u0026thinsp;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 significant association (P\u0026thinsp;=\u0026thinsp;0.064)). Those with experience of bloody urine accounted for the highest frequency [11.5% (15/131)] with significant association (P\u0026thinsp;=\u0026thinsp;0.017). Infected People with blood-brown urine colour had the highest percentage of the antigen [13/68 (19.1%)], with significant association (P\u0026thinsp;=\u0026thinsp;0.000).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFrequency of the Sh 22.6 Antigen Among the Infected Individuals for the Clinical Manifestations\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eClinical Manifestations\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. Examined\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNo. Positive\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePrevalence (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eX\u003csup\u003e2\u003c/sup\u003e-Value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eDF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eP-Value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDifficult Urination\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e154\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.000*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e127\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e12.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e149\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePainful Urine\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.843\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.064\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e221\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBloody Urine Experience\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e131\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e36.763\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.017*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e145\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePhysical Observation\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBlood Brown\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e19.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25.583\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.000*\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDark Brown\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLight Brown\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLight Yellow\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e157\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCom_B 11 Strip\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHaematuria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e43.761\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.083\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eProteinuria\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNormal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e276\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e6.2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e*= Significantly Associated; DF\u0026thinsp;=\u0026thinsp;Degree of Freedom (n-1)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThe present study on the detection of \u003cem\u003eS. haematobium\u003c/em\u003e antigen (Sh 22.6 kDa) showed that 6.2% of the infected individuals in the study area, were harbouring \u003cem\u003eS. haematobium\u003c/em\u003e 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., (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) made a similar observation while conducting an expression on analysis of highly polymorphic mucin proteins (Sm PoMuc) from the parasite of \u003cem\u003eSchistosoma species;\u003c/em\u003e 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% identified in studying increased ShTAL1 (sh 22.6kDa) against IgE before praziquantel treatment in Ghanaians\u0026rsquo; \u003cem\u003eS. haematobium\u003c/em\u003e endemic community (Asuming-Brempong et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Also, it was lower than the 35.5% identified by Fitzsimmons et al. (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) while investigating human IgE response to the \u003cem\u003eS. haematobium\u003c/em\u003e 22\u0026middot;6 kDa antigen. However, it was in the same range with 10.3% identified in Gabon by Webster \u003cem\u003eet al.\u003c/em\u003e (2016) in their research on human immunoglobulin E responses to a recombinant 22.6-kilodalton antigen from \u003cem\u003eSchistosoma\u003c/em\u003e adult worms are associated with low intensities of reinfection after treatment.\u003c/p\u003e \u003cp\u003eAge group is significantly 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 (\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;56 years old) to produce a high amount of IgE which responds to a 22.6-kilodalton antigen from \u003cem\u003eSchistosoma\u003c/em\u003e adult worms. The present finding was consistent with earlier research in communities with endemic \u003cem\u003eS. mansoni\u003c/em\u003e and \u003cem\u003eS. haematobium\u003c/em\u003e infections by Dunne et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), 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. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), on human IgE, IgG4 and resistance to reinfection with \u003cem\u003eS. haematobium\u003c/em\u003e also showed that higher production of the resistance antigen among the \u003cem\u003eS. haematobium\u003c/em\u003e infected is associated with older age. The current finding did not line up with earlier research by Dunne et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), on the isolation of a 22 kDa band after SDS-PAGE of \u003cem\u003eSchistosoma\u003c/em\u003e 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 \u003cem\u003eSchistosoma\u003c/em\u003e infections whose IgE antibodies recognized antigen(s) in the 22-kDa range were less infected with the antigen.\u003c/p\u003e \u003cp\u003eGender was significantly 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\u0026rsquo; exposure to the parasites, which may lead to increased antigen production. A similar finding was made in an investigation of people who had \u003cem\u003eS. japonicum\u003c/em\u003e infection in the Philippines (Webster et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). 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 \u003cem\u003eet al.\u003c/em\u003e, 1997), and the development of gender-specific immunity is supported by the findings of the current investigation.\u003c/p\u003e \u003cp\u003eClinical manifestations such as bloody urine experience and blood brown urine are significantly associated with the prevalence of Sh 22.6 kDa antigen in the study area. This could be attributed to the fact that infection with \u003cem\u003eS. haematobium\u003c/em\u003e affects the seminal vesicles, lower ureters, and bladder, the oxygen gradient created by the urine fluid 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 Dvorǎ et al. (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Dunne et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), 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. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) made similar observations.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe present study showed that the antigen (Sh 22.6 kDa) started emerging among the \u003cem\u003eS. haematobium-\u003c/em\u003einfected people in the study area, however, currently, a small proportion of the people (6.2%) harboured the antigen. The frequency of the antigen was significantly 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.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eSh 22.6 kDa = \u003cem\u003eSchistosoma haematobium\u003c/em\u003e 22.6 kilodalton antigen\u003c/p\u003e\n\u003cp\u003eSDS-PAGE = Sodium Dodecyl Sulphate- Polyacrylamide Gel Electrophoreses\u003c/p\u003e\n\u003cp\u003eWB= Western Blotting\u003c/p\u003e\n\u003cp\u003eDNA=Deoxy Ribonucleic Acid\u003c/p\u003e\n\u003cp\u003eIgE= Immunoglobulin E\u003c/p\u003e\n\u003cp\u003eDF= Degree of Freedom\u003c/p\u003e\n\u003cp\u003eX\u003csup\u003e2\u003c/sup\u003e= Chi-Square\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics Approval and Consent to Participants\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe 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 (SKHREC/10/2023). When seeking consent from each individual, the principles, aim, objectives procedure, and significance of the study were explained to the participants using the local language (Hausa), and\u0026nbsp;individual(s) who were unwilling and those who either refused to return samples were not included in the survey.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data Set and Material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declared no competing interest\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo funds have been received for this research\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSJ 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.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe 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.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAfifi, M., Jiman-Fatani, A., Al-Hussainy, N., Al-Rabia, M. \u0026amp; Bogari, A. (2016). Genetic diversity among natural populations of \u003cem\u003eSchistosoma haematobium\u003c/em\u003e might contribute to inconsistent virulence and diverse clinical outcomes. \u003cem\u003eJournal of Microscopic Ultrastructures\u003c/em\u003e, \u003cstrong\u003e4\u003c/strong\u003e(4): 222-229. https://doi.org/10.1038/s41598-022-11645-3\u003c/li\u003e\n\u003cli\u003eAgi, P. I., \u0026amp; Okafor, E. J. (2005). The epidemiology of \u003cem\u003eSchistosoma haematobium\u003c/em\u003e in Odau community in the Niger Delta Area of Nigeria. \u003cem\u003eJournal of Applied Science and Environmental Management\u003c/em\u003e. \u003cstrong\u003e9\u003c/strong\u003e(3): 37-43.\u003c/li\u003e\n\u003cli\u003eArshad, M. \u0026amp; Odeh, G. O. (2014). The Establishment of the Nigerian Sokoto Caliphate: An inquest into the Background History of the 1804 Jihad in Hausa Land, 210 years After, \u003cem\u003eAl-Qalam\u003c/em\u003e \u003cem\u003eJournal of Arts and Social Sciences,\u003c/em\u003e \u003cstrong\u003e2\u003c/strong\u003e(1): 61-79\u003c/li\u003e\n\u003cli\u003eAsmaa, M. El-Kady, M., EL-Amir, M. H. H., Khaled, S. A., Ahmad, A. \u0026amp; Alzahraa, A. A. (2020). Genetic Diversity of \u003cem\u003eSchistosoma haematobium \u003c/em\u003ein Qena Governorate, Upper Egypt.\u003cem\u003e Infection and Drug Resistance, \u003c/em\u003e\u003cstrong\u003e13\u003c/strong\u003e(2): 3601\u0026ndash;3611\u003c/li\u003e\n\u003cli\u003eAsuming-Brempong, E. K., Ayi, I., van der Puije, W., Gyan, B. A., Larbi, I. A., Ashong, Y., Frempong, N. A., Quartey, J. K., Otchere, J., Jones, F. M., Wilson, S., Dunne, D. W., \u0026amp; Boakye, D. A. (2022). Increased ShTAL1 IgE responses post-Praziquantel treatment may be associated with a reduced risk of re-infection in a Ghanaian S. haematobium-endemic community. \u003cem\u003ePLoS Neglected Tropical Diseases\u003c/em\u003e, \u003cem\u003e16\u003c/em\u003e(3). https://doi.org/10.1371/journal.pntd.0010115\u003c/li\u003e\n\u003cli\u003eAtalabi, T. E., Lawal, U., \u0026amp; Ipinlaye, S. J. (2016). Prevalence and intensity of genito-urinary schistosomiasis and associated risk factors among junior high school students in two local government areas around Zobe Dam in Katsina State, Nigeria. \u003cem\u003eParasites and Vectors\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(1). https://doi.org/10.1186/s13071-016-1672-5\u003c/li\u003e\n\u003cli\u003eBlack, C. L., Mwinzi, P. N. M., Muok, E. M. O., Abudho, B., Fitzsimmons, C. M., Dunne, D. W., Karanja, D. M. S., Secor, W. E., \u0026amp; Colley, D. G. (2010). Influence of exposure history on the immunology and development of resistance to human Schistosomiasis mansoni. \u003cem\u003ePLoS Neglected Tropical Diseases\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(3). https://doi.org/10.1371/journal.pntd.0000637\u003c/li\u003e\n\u003cli\u003eBosompem, K. M., Arishima, T., Yamashita, T., Ayi, I., Anyan, W. K., \u0026amp; Kojima, S. (1996). Extraction of \u003cem\u003eSchistosoma haematobium\u003c/em\u003e antigens from infected human urine and generation of potential diagnostic monoclonal antibodies to urinary antigens\u003cem\u003e. Acta Tropica, \u003c/em\u003e\u003cstrong\u003e62\u003c/strong\u003e(2), 91\u0026ndash;103\u003cem\u003e.\u003c/em\u003e doi:10.1016/s0001-706x(96)00040-x \u003c/li\u003e\n\u003cli\u003eCasacuberta-Partal, M., Janse, J. J., van Schuijlenburg, R., de Vries, J. J. C., Erkens, M. A. A., Suijk, K., van Aalst, M., Maas, J. J., Grobusch, M. P., van Genderen, P. J. J., de Dood, C., Corstjens, P. L. A. M., van Dam, G. J., van Lieshout, L., \u0026amp; Roestenberg, M. (2021). Antigen-based diagnosis of Schistosoma infection in travellers: A prospective study. \u003cem\u003eJournal of Travel Medicine\u003c/em\u003e, \u003cem\u003e27\u003c/em\u003e(4). https://doi.org/10.1093/JTM/TAAA055\u003c/li\u003e\n\u003cli\u003eChristine, P., Shona, W., He\u0026acute;le`ne, L., Sophia, L., Frances, M. J., Wassim, D., Monique, C., David, W. D. \u0026amp; Jamal, K. (2006). Identification of a Novel Antigen of \u003cem\u003eSchistosoma mansoni\u003c/em\u003e Shared with Plasmodium falciparum and Evaluation of Different Cross-Reactive Antibody Subclasses Induced by Human Schistosomiasis and Malaria. \u003cem\u003eInfection and Immunity\u003c/em\u003e, \u003cstrong\u003e74\u003c/strong\u003e(6) 3347\u0026ndash;3354. \u003c/li\u003e\n\u003cli\u003eDaumerie, Denis., Savioli, Lorenzo., Crompton, D. W. T. (David W. T., \u0026amp; World Health Organization. Department of Control of Neglected Tropical Diseases. (2010). \u003cem\u003eWorking to overcome the global impact of neglected tropical diseases: first WHO report on neglected tropical diseases.\u003c/em\u003e World Health Organization\u003c/li\u003e\n\u003cli\u003eDi Bella, S., Riccardi, N., Giacobbe, D. R., \u0026amp; Luzzati, R. (2018). History of schistosomiasis (bilharziasis) in humans: from Egyptian medical papyri to molecular biology on mummies. \u003cem\u003ePathogens and Global Health\u003c/em\u003e, \u003cem\u003e112\u003c/em\u003e(5), 268\u0026ndash;273. https://doi.org/10.1080/20477724.2018.1495357\u003c/li\u003e\n\u003cli\u003eDunne, D. W., Butterworth, A. E., \u0026amp; Fulford, A. J. (2017). Immunity after treatment of human schistosomiasis: association between IgE antibodies to adult worm antigens and resistance to reinfection. \u003cem\u003eEuropean Journal Immunology.\u003c/em\u003e \u003cstrong\u003e22\u003c/strong\u003e: 1483\u0026ndash;1494.\u003c/li\u003e\n\u003cli\u003eDunne, D. W., Webster, M., Smith, P., Langley, J. G., Richardson, B. A., Fulford, A. J. C., Butterworth, A. E., Sturrock, R. F., Kariuki, H. C. \u0026amp; Ouma, J. H. (2021): The isolation of a 22 kDa band after SDS-PAGE of \u003cem\u003eSchistosoma mansoni\u003c/em\u003e adult worms and its use to demonstrate that IgE responses against the antigen(s) it contains are associated with human resistance to reinfection. \u003cem\u003eParasite Immunology. \u003c/em\u003e19:79\u0026ndash;89.\u003c/li\u003e\n\u003cli\u003eDvorǎ, ́K. J., Mashiyama, S. T., Braschi, S., Sajid, M., Knudsen, G. M., \u0026amp; Hansell, E, (2018). Differential use of protease families for invasion by schistosome cercariae.\u003cem\u003e Biochemistry.\u003c/em\u003e \u003cstrong\u003e90\u003c/strong\u003e(2):345\u0026ndash;358\u003c/li\u003e\n\u003cli\u003eEl-Kady, A. M., El-Amir, M. I., Hassan, M. H., Allemailem, K. S., Almatroudi, A., \u0026amp; Ahmad, A. A. (2020). Genetic diversity of \u003cem\u003eSchistosoma haematobium\u003c/em\u003e in Qena governorate, Upper Egypt. \u003cem\u003eInfection and Drug Resistance\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e, 3601\u0026ndash;3611. https://doi.org/10.2147/IDR.S266928\u003c/li\u003e\n\u003cli\u003eEl-Kady, A. M., El-Amir, M. I., Hassan, M. H., Allemailem, K. S., Almatroudi, A., \u0026amp; Ahmad, A. A. (2020). Genetic diversity of \u003cem\u003eSchistosoma haematobium\u003c/em\u003e in Qena governorate, Upper Egypt. \u003cem\u003eInfection and Drug Resistance\u003c/em\u003e, \u003cem\u003e13\u003c/em\u003e, 3601\u0026ndash;3611. https://doi.org/10.2147/IDR.S266928\u003c/li\u003e\n\u003cli\u003eFitzsimmons, C. M., Stewart, T. J., Hoffmann, K. F., Grogan, J. L., Yazdanbakhsh, M., \u0026amp; Dunne, D. W. (2004). Human IgE response to the \u003cem\u003eSchistosoma haematobium\u003c/em\u003e 22\u0026middot;6 kDa antigen. In \u003cem\u003eParasite Immunology\u003c/em\u003e (Vol. 26). www.sanger.ac.uk\u003c/li\u003e\n\u003cli\u003eGalazka, A. M. (1993). \u003cem\u003eThe Immunological Basis for Immunization Series General Immunology Medical Officer Expanded Programme on Immunization global programme for vaccines and immunization expanded programme on immunization The Expanded Programme on Immunization thanks the following donors whose support made the production of these modules possible: United Nations Development Fund (UNDP) The Rockefeller Foundation\u003c/em\u003e. http://www.who.ch/programmes/gpv/gEnglish/avail/gpvcatalog/catlog1.htm\u003c/li\u003e\n\u003cli\u003eGamde, S. M., Garba, A., Tongvwam, P., Gamde, S. D., Agom, D. D., \u0026amp; Igbinosa, I. (2023). Prevalence, intensity, and risk factors of urinary schistosomiasis among primary school children in Silame, Sokoto, Nigeria. \u003cem\u003eMicrobes and Infectious Diseases\u003c/em\u003e, \u003cem\u003e4\u003c/em\u003e(2), 695\u0026ndash;703. https://doi.org/10.21608/MID.2022.155035.1365\u003c/li\u003e\n\u003cli\u003eHagan P, Blumenthal UJ, Dunne D, Simpson AJG, Wilkins HA (2015): Human IgE, IgG4 and resistance to reinfection with \u003cem\u003eSchistosoma haematobium\u003c/em\u003e. \u003cem\u003eNature\u003c/em\u003e; \u003cstrong\u003e349\u003c/strong\u003e:243\u0026ndash;245. \u003c/li\u003e\n\u003cli\u003eHalligan, B.D., Rootti, V., Jin, W., Laffoo, S., Twigger, S.N., \u0026amp; Dratz, E.A., (2004). ProMoSt [protein Modification screening tool] is a web-based tool for mapping protein modification of two-dimensional gels. \u003cem\u003eNucleic Acids Research,\u003c/em\u003e \u003cstrong\u003e32:\u003c/strong\u003e638-644.\u003c/li\u003e\n\u003cli\u003eHamdan, M., \u0026amp; Righetti, P. G., (2005). John Wiley \u0026amp; Sons, Inc., Proteomics Today: Protein Assessment and Biomarkers Using Mass Spectrometry, Electrophoresis, and Microarray Technology, \u003cem\u003eHoboken, Nature Journal, USA.\u003c/em\u003e 274\u0026ndash;340.\u003c/li\u003e\n\u003cli\u003eHany Sady, Hesham M. Al-Mekhlafi, Romano Ngui, Wahib M. Atroosh, Ahmed K. Al-Delaimy, Nabil A. Nasr, Salwa Dawaki, Awatif M. Abdulsalam, Init Ithoi, Yvonne A. L. Lim, Kek Heng Chua, \u0026amp; Johari, S. (2015). Detection of \u003cem\u003eSchistosoma mansoni\u003c/em\u003e and \u003cem\u003eSchistosoma haematobium\u003c/em\u003e by PCR with High-Resolution Melting Analysis \u003cem\u003eInternational Journal of Molecular Science.\u003c/em\u003e \u003cstrong\u003e16\u003c/strong\u003e, 16085-16103\u003c/li\u003e\n\u003cli\u003eIsmail, A., \u0026amp; Oke, I. (2012). Statistical and trend analyses of rainfall in Sokoto. \u003cem\u003eInternational Research Journal of Engineering Science, Technology and Innovation\u003c/em\u003e, \u003cstrong\u003e1\u003c/strong\u003e: 161\u0026ndash;174\u003c/li\u003e\n\u003cli\u003eJibrillah, A. M., Ja\u0026rsquo;afar, M., \u0026amp; Choy, L. K. (2019). Monitoring Vegetation Change in the Dryland Ecosystem of Sokoto, Northwestern Nigeria using Geoinformatics. \u003cem\u003eIndonesian Journal of Geography\u003c/em\u003e, \u003cstrong\u003e51\u003c/strong\u003e(1): 9\u0026ndash;17.\u003c/li\u003e\n\u003cli\u003eJiya, F. B., Jiya, N. M., Ibitoye, P. K., Mohammed, Y., \u0026amp; Mohammad, A. U. (2022). Chronic Urogenital Schistosomiasis among In-School Adolescents in Sokoto, North-Western Nigeria. \u003cem\u003eAsian Journal of Medicine and Health\u003c/em\u003e, 22\u0026ndash;36. https://doi.org/10.9734/ajmah/2022/v20i130429\u003c/li\u003e\n\u003cli\u003eKanamura, H. Y., Hoshino-Shimizu, S., Camargo, M. E. \u0026amp; Silva, L. C. (2019). Class-specific antibodies and fluorescent staining patterns in acute and chronic forms of \u003cem\u003eSchistosoma mansoni. American Journal and Tropical Medicine and Hygiene\u003c/em\u003e; 28: 242-248\u003c/li\u003e\n\u003cli\u003eKasim, I. \u0026amp; Usman, A. Y. (2021). Trend analysis of Temperature and Precipitation Change in Sokoto State, \u003cem\u003eNigeria Journal of Agricultural Economics and Rural Development.\u003c/em\u003e \u003cstrong\u003e7\u003c/strong\u003e(1): 941-951\u003c/li\u003e\n\u003cli\u003eLadan, M. U., Abubakar, U., Abdullahi, K., Bunza, M. D. A., Ladan, M. J., \u0026amp; Adamu, T. (2012). Studies on Urinary Schistosomiasis in Selected Villages around Gusau Dam Site, Zamfara State, Nigeria. \u003cem\u003eNigerian Journal of Basic and Applied Science\u003c/em\u003e, \u003cem\u003e20\u003c/em\u003e(3), 189\u0026ndash;194. http://www.ajol.info/index.php/njbas/index\u003c/li\u003e\n\u003cli\u003eLodh N, Naples, J. M., Bosompem, K. M., Quartey, J. \u0026amp; Shiff, C. J. (2014). Detection of Parasite-Specific DNA in Urine Sediment Obtained by Filtration Differentiates between Single and Mixed Infections of \u003cem\u003eSchistosoma mansoni\u003c/em\u003e and\u003cem\u003e S. haematobium \u003c/em\u003efrom Endemic Areas in Ghana. \u003cem\u003ePLoS ONE. \u003c/em\u003e \u003cstrong\u003e9\u003c/strong\u003e(3): e91144. \u003c/li\u003e\n\u003cli\u003eMarkakpo, U. S., Armah, G. E., Fobil, J. N., Asmah, R. H., Anim-Baidoo, I., Dodoo, A. K., Madjitey, P., Essuman, E. E., Kojima, S., \u0026amp; Bosompem, K. M. (2015). Immunolocalization of the 29 kDa \u003cem\u003eSchistosoma haematobium\u003c/em\u003e species-specific antigen: A potential diagnostic marker for urinary schistosomiasis. \u003cem\u003eBMC Infectious Diseases\u003c/em\u003e, \u003cem\u003e15\u003c/em\u003e(1). https://doi.org/10.1186/s12879-015-0931-y\u003c/li\u003e\n\u003cli\u003eMohammed, I. H., Mohammed, H. I., Yahaya, I., \u0026amp; Buhari, S. B. (2020). Prevalence and Determining Factors of Urinary Schistosomiasis among Primary School Pupils in North Central Nigeria. In \u003cem\u003eAsian Journal of Parasitology\u003c/em\u003e (Vol. 4, Issue 3).\u003c/li\u003e\n\u003cli\u003eMwai, J., Njenga, S., \u0026amp; Barasa, M. (2016). Knowledge, attitude and practices concerning prevention and control of schistosomiasis infection in Mwea Kirinyaga county, Kenya. In \u003cem\u003eBMC Public Health\u003c/em\u003e (Vol. 16, Issue 1). BioMed Central Ltd. https://doi.org/10.1186/s12889-016-3494-y\u003c/li\u003e\n\u003cli\u003eOlds GR, Olveda R, Wu G, Wiest P, McGarvey S, Aligui G, Zhang S, Ramirez B, Daniel B, Peters P, Romulo R, Fevidal P, Tiu W, Yuan J, Domingo E. \u0026amp; Blas B (1996): Immunity and morbidity in schistosomiasis japonicum infection.\u003cem\u003e American Journal of Tropical Medicine Hygiene\u003c/em\u003e; \u003cstrong\u003e55\u003c/strong\u003e(5):121\u0026ndash;126.\u003c/li\u003e\n\u003cli\u003ePearson, M. S., Tedla, B. A., Mekonnen, G. G., Proietti, C., Becker, L., Nakajima, R., Jasinskas, A., Doolan, D. L., Amoah, A. S., Knopp, S., Rollinson, D., Ali, S. M., Kabole, F., Hokke, C. H., Adegnika, A. A., Field, M. A., van Dam, G., Corstjens, P. L. A. M., Mduluza, T. \u0026amp; Loukas, A. (2021). Immunomics-guided discovery of serum and urine antibodies for diagnosing urogenital schistosomiasis: a biomarker identification study. \u003cem\u003eThe Lancet Microbe\u003c/em\u003e, \u003cem\u003e2\u003c/em\u003e(11), e617\u0026ndash;e626. https://doi.org/10.1016/S2666-5247(21)00150-6\u003c/li\u003e\n\u003cli\u003ePinto-Almeida, A., Mendes, T., de Oliveira, R. N., Corr\u0026ecirc;a, S. de A. P., Allegretti, S. M., Belo, S., Tom\u0026aacute;s, A., Anibal, F. de F., Carrilho, E., \u0026amp; Afonso, A. (2016). Morphological characteristics of \u003cem\u003eSchistosoma mansoni\u003c/em\u003e PZQ-resistant and -susceptible strains are different in the presence of Praziquantel. \u003cem\u003eFrontiers in Microbiology\u003c/em\u003e, \u003cem\u003e7\u003c/em\u003e(APR). https://doi.org/10.3389/fmicb.2016.00594\u003c/li\u003e\n\u003cli\u003eRoger E, Gourbal B, Grunau C, Pierce RJ, Galinier R, \u0026amp; Mitta G. (2017). Expression analysis of highly polymorphic mucin proteins (Sm PoMuc) from the parasite \u003cem\u003eSchistosoma mansoni. Molecular Biochemistry and Parasitology.\u003c/em\u003e \u003cstrong\u003e157\u003c/strong\u003e(2):217\u0026ndash; 27.\u003c/li\u003e\n\u003cli\u003eSantiago, M. L. Hafalla, J. C. \u0026amp; Kurtis J. D. (2018)., \u0026ldquo;Identification of the \u003cem\u003eSchistosoma japonicum\u003c/em\u003e 22.6-kDa antigen as a major target of the human IgE response: similarity of IgE-binding epitopes to allergen peptides,\u0026rdquo; \u003cem\u003eInternational Archives of Allergy and Immunology\u003c/em\u003e, \u003cstrong\u003e117(\u003c/strong\u003e2): 94\u0026ndash;104.\u003c/li\u003e\n\u003cli\u003eSantos, L. L., Santos, J., Gouveia, M. J., Bernardo, C., Lopes, C., Rinaldi, G., Brindley, P. J., \u0026amp; da Costa, J. M. C. (2021). Review urogenital schistosomiasis\u0026mdash;history, pathogenesis, and bladder cancer. In \u003cem\u003eJournal of Clinical Medicine\u003c/em\u003e (Vol. 10, Issue 2, pp. 1\u0026ndash;11). MDPI. https://doi.org/10.3390/jcm10020205\u003c/li\u003e\n\u003cli\u003eSenghor, B., Diaw, O. T., Doucoure, S., Sylla, S. N., Seye, M., Talla, I., B\u0026acirc;, C. T., Diallo, A., \u0026amp; Sokhna, C. (2015). Efficacy of praziquantel against urinary schistosomiasis and reinfection in Senegalese school children where there is a single well-defined transmission period. \u003cem\u003eParasites and Vectors\u003c/em\u003e, \u003cem\u003e8\u003c/em\u003e(1). https://doi.org/10.1186/s13071-015-0980-5\u003c/li\u003e\n\u003cli\u003eSepulveda, J., Tremblay, J. M., DeGnore, J. P., Skelly, P. J., \u0026amp; Shoemaker, C. B. (2010). \u003cem\u003eSchistosoma mansoni\u003c/em\u003e host-exposed surface antigens characterized by sera and recombinant antibodies from schistosomiasis-resistant rats. \u003cem\u003eInternational Journal for Parasitology\u003c/em\u003e, \u003cem\u003e40\u003c/em\u003e(12), 1407\u0026ndash;1417. https://doi.org/10.1016/j.ijpara.2010.04.019\u003c/li\u003e\n\u003cli\u003eSingh, K., Muddasiru, D., \u0026amp; Singh, J. (2016). Current status of schistosomiasis in Sokoto, Nigeria. \u003cem\u003eParasite Epidemiology and Control\u003c/em\u003e, \u003cem\u003e1\u003c/em\u003e(3), 239\u0026ndash;244. https://doi.org/10.1016/j.parepi.2016.08.003\u003c/li\u003e\n\u003cli\u003eSuleiman, J., T. Isyaku, N., E. Ukatu, V., \u0026amp; Alhaji, Y. I. (2022). Antigens of Three Medically Important Schistosoma spp. \u003cem\u003eAsian Journal of Research in Zoology\u003c/em\u003e, 11\u0026ndash;27. https://doi.org/10.9734/ajriz/2022/v5i330139\u003c/li\u003e\n\u003cli\u003eTowbin, H., Staehlin, T. and Gordon, J. (2019). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. \u003cem\u003eProcedures for National Academic Science, \u003c/em\u003e\u003cstrong\u003e76\u003c/strong\u003e\u003cem\u003e: \u003c/em\u003e4350\u0026ndash;4354.\u003c/li\u003e\n\u003cli\u003eVale, N., Gouveia, M. J., Rinaldi, G., Brindley, P. J., G\u0026auml;rtner, F., \u0026amp; Da Costa, J. M. C. (2017). Praziquantel for schistosomiasis: Single-drug metabolism revisited, mode of action, and resistance. In \u003cem\u003eAntimicrobial Agents and Chemotherapy\u003c/em\u003e (Vol. 61, Issue 5). American Society for Microbiology. https://doi.org/10.1128/AAC.02582-16\u003c/li\u003e\n\u003cli\u003eVinkeles, N., Melchers, V. S., van Dam, G. J., Shaproski, D., Kahama, A. I., Brienen, E. A. T., Vennervald, B. J., \u0026amp; van Lieshout, L. (2014). Diagnostic Performance of Schistosoma Real-Time PCR in Urine Samples from Kenyan Children Infected with \u003cem\u003eSchistosoma haematobium\u003c/em\u003e: Day-to-day Variation and Follow-up after Praziquantel Treatment. \u003cem\u003ePLoS Neglected Tropical Diseases\u003c/em\u003e, \u003cstrong\u003e8\u003c/strong\u003e(4). https://doi.org/10.1371/journal.pntd.0002807\u003c/li\u003e\n\u003cli\u003eWebster, M., Ramirez, B. L., Aligui, G. D., Olveda, R. M., Ouma, J. H., Kariuki, H. C., Kimani, G., Olds, G. R., Fulford, J. C., Butterworth, A. E. \u0026amp; Dunne, D. W. (1997). The influence of sex and age on antibody isotype responses to \u003cem\u003eSchistosoma mansoni\u003c/em\u003e and \u003cem\u003eSchistosoma japonicum\u003c/em\u003e in human populations in Kenya and the Philippines. \u003cem\u003eParasitology\u003c/em\u003e; 114:383\u0026ndash;393\u003c/li\u003e\n\u003cli\u003eWorld Health Organization (2002). Prevention and control of schistosomiasis and soil-transmitted helminthiasis. 2003/02/21 ed. \u003cem\u003eGeneva: World Health Organization\u003c/em\u003e; i\u0026ndash;vi, 1\u0026ndash;57, back cover p\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-parasitic-diseases","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jopd","sideBox":"Learn more about [Journal of Parasitic Diseases](https://www.springer.com/journal/12639)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/jopd/default.aspx","title":"Journal of Parasitic Diseases","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Antigen, Emergence, Filtration Technique, PCR, Riverine Communities, Schistosoma haematobium, SDS-PAGE, Urine","lastPublishedDoi":"10.21203/rs.3.rs-4843645/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4843645/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eA cross-sectional study was conducted to investigate the emergence of Sh 22.6 kDa antigen among the \u003cem\u003eS. haematobium\u003c/em\u003e 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 filtration method, all microscopy-positive samples were subjected to PCR analysis for confirmation 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.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eOut of 276 PCR-positive individuals, 6.2% (17/276) were harbouring Sh 22.6 kDa antigen. The distribution of the antigen was significantly associated with the location (P\u0026thinsp;\u0026gt;\u0026thinsp;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 significant association (P\u0026thinsp;=\u0026thinsp;0.000). Although, the age group is not significantly associated with the antigen frequency (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05), infected people aged\u0026thinsp;\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e\u0026ge;\u003c/span\u003e\u0026thinsp;56 years old had the highest Sh 22.6 kDa antigen [40.0% (2/5)] than other age groups. Bloody urine experience (P\u0026thinsp;=\u0026thinsp;0.017) and urine colour (P\u0026thinsp;=\u0026thinsp;0.000) were the study area's clinical manifestations associated with Sh 22.6 kDa distribution.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThe present study showed that a small proportion of the infected people harboured \u003cem\u003eS. haematobium\u003c/em\u003e which has Sh 22.6 kDa antigen. Therefore, health education campaigns and mass chemotherapy should be considered by the authorities concerned in the fight against urinary schistosomiasis in the area.\u003c/p\u003e","manuscriptTitle":"Emergence of Sh 22.6 kDa Antigen Among the Schistosoma haematobium Infected People in Riverine Communities of the Extreme North Western Nigeria","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-30 05:24:24","doi":"10.21203/rs.3.rs-4843645/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Minor revisions needed","date":"2025-07-17T08:08:02+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2025-06-09T11:37:03+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-08-05T10:12:06+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-08-02T08:29:51+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Parasitic Diseases","date":"2024-08-01T12:35:08+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-parasitic-diseases","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jopd","sideBox":"Learn more about [Journal of Parasitic Diseases](https://www.springer.com/journal/12639)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/jopd/default.aspx","title":"Journal of Parasitic Diseases","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"11aef642-6584-42d7-8674-1098fbf79d67","owner":[],"postedDate":"August 30th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-08-13T06:48:24+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-30 05:24:24","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4843645","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4843645","identity":"rs-4843645","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}