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PLoS Neglected Tropical Diseases Mar 2021We were tasked by the World Health Organization (WHO) to address the following question: What techniques should be used to diagnose Schistosoma infections in snails and...
BACKGROUND
We were tasked by the World Health Organization (WHO) to address the following question: What techniques should be used to diagnose Schistosoma infections in snails and in the water in potential transmission sites? Our goal was to review and evaluate the available literature and provide recommendations and insights for the development of WHO's Guidelines Development Group for schistosomiasis control and elimination.
METHODOLOGY
We searched several databases using strings of search terms, searched bibliographies of pertinent papers, and contacted investigators who have made contributions to this field. Our search covered from 1970 to Sept 2020. All papers were considered in a PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) framework, and retained papers were grouped by technique and subjected to our GRADE (Grading of Recommendations, Assessment, Development and Evaluations) evidence assessment profile determined in consultation with WHO. We also considered issues of sensitivity, specificity, coverage, cost, robustness, support needs, schistosome species discrimination, and relevant detection limits.
PRINCIPAL FINDINGS
Our PRISMA process began with the perusal of 949 articles, of which 158 were retained for data extraction and evaluation. We identified 25 different techniques and for each applied a GRADE assessment considering limitations, inconsistency, imprecision, indirectness, and publication bias. We also provide advantages and disadvantages for each category of techniques.
CONCLUSIONS
Our GRADE analysis returned an assessment of moderate quality of evidence for environmental DNA (eDNA), qPCR and LAMP (Loop-mediated isothermal amplification). No single ideal diagnostic approach has yet been developed, but considerable recent progress has been made. We note a growing trend to use eDNA techniques to permit more efficient and replicable sampling. qPCR-based protocols for follow-up detection offer a versatile, mature, sensitive, and specific platform for diagnosis though centralized facilities will be required to favor standardization. Droplet digital PCR (ddPCR) can play a complementary role if inhibitors are a concern, or more sensitivity or quantification is needed. Snail collection, followed by shedding, is encouraged to provide specimens for sequence verifications of snails or schistosomes. LAMP or other isothermal detection techniques offer the prospect of less expensive and more distributed network of analysis but may face standardization and verification challenges related to actual sequences amplified. Ability to detect schistosome infections in snails or in the water is needed if control and elimination programs hope to succeed. Any diagnostic techniques used need to be regularly verified by the acquisition of DNA sequences to confirm that the detected targets are of the expected species. Further improvements may be necessary to identify the ideal schistosome or snail sequences to target for amplification. More field testing and standardization will be essential for long-term success.
Topics: Animals; DNA, Environmental; DNA, Helminth; Molecular Diagnostic Techniques; Nucleic Acid Amplification Techniques; Real-Time Polymerase Chain Reaction; Schistosoma; Schistosomiasis; Snails; Water
PubMed: 33760814
DOI: 10.1371/journal.pntd.0009175 -
PLoS Neglected Tropical Diseases Mar 2021The antihelminthic drug praziquantel has been used as the drug of choice for treating schistosome infection for more than 40 years. Although some epidemiological studies... (Meta-Analysis)
Meta-Analysis
BACKGROUND
The antihelminthic drug praziquantel has been used as the drug of choice for treating schistosome infection for more than 40 years. Although some epidemiological studies have reported low praziquantel efficacy in cure rate (CR) and/or egg reduction rate (ERR), there is no consistent robust evidence of the development of schistosome resistance to praziquantel (PZQ). There is need to determine factors that lead to variable treatment CR and/or ERR. Therefore, we conducted a systematic review and meta-analysis to review CR and ERR as well as identify their predictors.
METHODOLOGY/PRINCIPAL FINDINGS
In this systematic review and meta-analysis, a literature review was conducted using Biosis Citation Index, Data Citation Index, MEDLINE, and Web of Science Core Collection all of which were provided through Web of Science. Alongside these, EMBASE, and CAB abstracts were searched to identify relevant articles. Random effect meta-regression models were used to identify the factors that influence CR and/or ERR by considering differences in host characteristics and drug dose. In total, 12,127 potential articles were screened and 146 eligible articles (published from 1979 to 2020) were identified and included for the meta-analysis. We found that there has been no significant reduction in CR or ERR over the study period. The results showed more variability in CR, compared with ERR which was more consistent and remained high. The results showed a positive effect of "PZQ treatment dose" with the current recommended dose of 40 mg/kg body weight achieving 57% to 88% CR depending on schistosome species, age of participants, and number of parasitological samples used for diagnosis, and ERR of 95%.
CONCLUSIONS/SIGNIFICANCE
Based on a review of over 40 years of research there is no evidence to support concerns about schistosomes developing resistance to PZQ. These results indicate that PZQ remains effective in treating schistosomiasis.
Topics: Animals; Anthelmintics; Humans; Parasite Egg Count; Praziquantel; Schistosoma; Schistosomiasis; Treatment Outcome
PubMed: 33730095
DOI: 10.1371/journal.pntd.0009189 -
PLoS Neglected Tropical Diseases Mar 2021Most of national schistosomiasis elimination programmes in Asia are relying on stool examination, particularly Kato Katz stool examination technique for regular... (Meta-Analysis)
Meta-Analysis
BACKGROUND
Most of national schistosomiasis elimination programmes in Asia are relying on stool examination, particularly Kato Katz stool examination technique for regular transmission monitoring. However, the Kato-Katz technique has shown low sensitivity for the detection of light-intensity infections, and therefore highly sensitive diagnostic tools are urgently required to monitor prevalence of infection in low transmission settings. The objective of this systematic review was to evaluate and synthesize the performance of diagnostic tests for detecting Schistosoma japonicum and S. mekongi infection in people living in endemic areas.
METHODOLOGY/PRINCIPAL FINDINGS
We comprehensively searched these nine electronic databases and other resources until July 2019, with no language or publication limits: PubMed, EMBASE, MEDLINE, Web of Science, BIOSIS Citation Index, HTA, CINAHL PLUS, The Cochrane Library, and PsycINFO. We included original studies that assessed diagnostic performance using antibody, antigen, and molecular tests with stool examination test as a reference standard. Two reviewers independently extracted a standard set of data and assessed study quality. We estimated the pooled estimates of sensitivity and specificity for each index test. We used diagnostic odds ratio to determine the overall accuracy and hierarchical summary receiver operating characteristics (HSROC) curve to assess the index tests performance. Fifteen studies (S. japonicum [n = 13] and S. mekongi [n = 2]) testing 15,303 participants were included in the review. Five studies reported performance of enzyme-linked immunosorbent assay (ELISA), seven studies reported indirect hemagglutination assay (IHA), and four studies reported polymerase chain reaction (PCR) for detecting S. japonicum. The pooled sensitivity and specificity were 0.93 (95% CI: 0.84-0.98) and 0.40 (95% CI: 0.29-0.53) for ELISA, 0.97 (95% CI: 0.90-0.99) and 0.66 (95% CI: 0.58-0.73) for IHA, and 0.89 (95% CI: 0.71-0.96) and 0.49 (95% CI: 0.29-0.69) for PCR respectively. A global summary indicated the best performance for IHA, closely followed by ELISA. We were unable to perform meta-analysis for S. mekongi due to insufficient number of studies.
CONCLUSIONS/SIGNIFICANCE
IHA showed the highest detection accuracy for S. japonicum. Further studies are needed to determine the suitable diagnostic methods to verify the absence of transmission of S. mekongi and also to compare detection accuracy against more sensitive reference standards such as PCR.
Topics: Animals; Enzyme-Linked Immunosorbent Assay; Fluorescent Antibody Technique, Indirect; Humans; Polymerase Chain Reaction; Schistosoma; Schistosoma japonicum
PubMed: 33730048
DOI: 10.1371/journal.pntd.0009244 -
Parasites & Vectors Mar 2021Schistosomiasis is a highly prevalent parasitic disease that can lead to adverse maternal and perinatal outcomes. To our knowledge, there has been no systematic review... (Meta-Analysis)
Meta-Analysis
BACKGROUND
Schistosomiasis is a highly prevalent parasitic disease that can lead to adverse maternal and perinatal outcomes. To our knowledge, there has been no systematic review and meta-analysis of schistosomiasis during pregnancy.
METHODS
We followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Relevant published studies were searched in international databases (PubMed, Science Direct, Scopus, Web of Science, and Google Scholar), from their inception until May 31, 2020. The retrieved studies were assessed for quality using the Modified Newcastle-Ottawa Scale. OpenMeta Analyst software was used for the statistical analysis.
RESULTS
Thirty-two studies enrolling 21024 pregnant women were included in this meta-analysis. All 32 of these studies were conducted in Africa. Of these studies, 19, 11, and 2 investigated S. mansoni, S. haematobium, and combined S. mansoni and S. haematobium infections, respectively. The pooled prevalence estimate of schistosomiasis during pregnancy was 13.2% (95 CI 11.0-15.4). A random model was used because of high heterogeneity (Q = 99.14; P < 0.001). In subgroup analyses, the pooled prevalence estimate of S. haematobium was significantly higher than the pooled prevalence estimates of S. mansoni [22.5% (95% CI 1.6-43.5) vs 8.7% (95% CI 6.0-11.3, P = 0.016), respectively]. The results of meta-regression analyses showed a non-significant difference in the prevalence of schistosomiasis during pregnancy according to the study sample sizes and year of publication. Only six studies evaluated the association between schistosomiasis during pregnancy and anemia. Schistosomiasis was associated with anemia in these six studies (OR = 3.02, 95% = 1.25‒7.28, P = 0.014).
CONCLUSION
The present meta-analysis suggests that schistosomiasis during pregnancy is an existing health problem. This meta-analysis also highlights the lack of data on the determinants and outcomes of schistosomiasis during pregnancy. Preventive measures are needed and could be part of antenatal care in areas endemic with schistosomiasis.
Topics: Africa; Anemia; Female; Humans; Pregnancy; Prevalence; Schistosomiasis; Schistosomiasis haematobia
PubMed: 33653391
DOI: 10.1186/s13071-021-04642-4 -
PLoS Neglected Tropical Diseases Feb 2021Schistosomiasis is a parasitic disease caused by trematode worms of the genus Schistosoma and belongs to the neglected tropical diseases. The disease has been reported...
BACKGROUND
Schistosomiasis is a parasitic disease caused by trematode worms of the genus Schistosoma and belongs to the neglected tropical diseases. The disease has been reported in 78 countries, with around 290.8 million people in need of treatment in 2018. Schistosomiasis is predominantly considered a rural disease with a subsequent focus of research and control activities in rural settings. Over the past decades, occurrence and even expansion of schistosomiasis foci in peri-urban and urban settings have increasingly been observed. Rural-urban migration in low- and middle-income countries and subsequent rapid and unplanned urbanization are thought to explain these observations. Fifty-five percent (55%) of the world population is already estimated to live in urban areas, with a projected increase to 68% by 2050. In light of rapid urbanization and the efforts to control morbidity and ultimately achieve elimination of schistosomiasis, it is important to deepen our understanding of the occurrence, prevalence, and transmission of schistosomiasis in urban and peri-urban settings. A systematic literature review looking at urban and peri-urban schistosomiasis was therefore carried out as a first step to address the research and mapping gap.
METHODOLOGY
Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a systematic computer-aided literature review was carried out using PubMed, ScienceDirect, and the World Health Organization Database in November 2019, which was updated in March 2020. Only papers for which at least the abstract was available in English were used. Relevant publications were screened, duplicates were removed, guidelines for eligibility were applied, and eligible studies were reviewed. Studies looking at human Schistosoma infections, prevalence, and intensity of infection in urban and peri-urban settings were included as well as those focusing on the intermediate host snails.
PRINCIPAL FINDINGS
A total of 248 publications met the inclusion criteria. The selected studies confirm that schistosomiasis is prevalent in peri-urban and urban areas in the countries assessed. Earlier studies report higher prevalence levels in urban settings compared to data extracted from more recent publications, yet the challenge of migration, rapid uncontrolled urbanization, and resulting poor living conditions highlight the potential for continuous or even newly established transmission to take place.
CONCLUSIONS
The review indicates that schistosomiasis has long existed in urban and peri-urban areas and remains a public health problem. There is, however, a challenge of comparability of settings due to the lack of a clear definition of what constitutes urban and peri-urban. There is a pressing need for improved monitoring of schistosomiasis in urban communities and consideration of treatment strategies.
Topics: Animals; Humans; Schistosoma; Schistosomiasis; Snails; Suburban Population; Urban Population
PubMed: 33630833
DOI: 10.1371/journal.pntd.0008995 -
PLoS Neglected Tropical Diseases Feb 2021Current knowledge on the burden of, and interactions between malaria and helminth co-infections, as well as the impact of the dual infections on anaemia, remains... (Meta-Analysis)
Meta-Analysis
BACKGROUND
Current knowledge on the burden of, and interactions between malaria and helminth co-infections, as well as the impact of the dual infections on anaemia, remains inconclusive. We have conducted a systematic review with meta-analysis to update current knowledge as a first step towards developing and deploying coordinated approaches to the control and, ultimately, elimination of malaria-helminth co-infections among children living in endemic countries.
METHODOLOGY/PRINCIPAL FINDINGS
We searched Medline, Embase, Global Health and Web of Science from each database inception until 16 March 2020, for peer-reviewed articles reporting malaria-helminth co-infections in children living in endemic countries. No language restriction was applied. Following removal of duplicates, two reviewers independently screened the studies for eligibility. We used the summary odds ratio (OR) and 95% confidence intervals (CI) as a measure of association (random-effects model). We also performed Chi-square heterogeneity test based on Cochrane's Q and evaluated the severity of heterogeneity using I2 statistics. The included studies were examined for publication bias using a funnel plot and statistical significance was assessed using Egger's test (bias if p<0.1). Fifty-five of the 3,507 citations screened were eligible, 28 of which had sufficient data for meta-analysis. The 28 studies enrolled 22, 114 children in 13 countries across sub-Saharan Africa, Southeast Asia and South America. Overall, the pooled estimates showed a prevalence of Plasmodium-helminth co-infections of 17.7% (95% CI 12.7-23.2%). Summary estimates from 14 studies showed a lower odds of P. falciparum infection in children co-infected with Schistosoma spp (OR: 0.65; 95%CI: 0.37-1.16). Similar lower odds of P. falciparum infection were observed from the summary estimates of 24 studies in children co-infected with soil transmitted helminths (STH) (OR: 0.42; 95%CI: 0.28-0.64). When adjusted for age, gender, socio-economic status, nutritional status and geographic location of the children, the risk of P. falciparum infection in children co-infected with STH was higher compared with children who did not have STH infection (OR = 1.3; 95% CI 1.03-1.65). A subset of 16 studies showed that the odds of anaemia were higher in children co-infected with Plasmodium and STH than in children with Plasmodium infection alone (OR = 1.20; 95% CI: 0.59-2.45), and were almost equal in children co-infected with Plasmodium-Schistosoma spp or Plasmodium infection alone (OR = 0.97, 95% CI: 0.30-3.14).
CONCLUSIONS/SIGNIFICANCE
The current review suggests that prevalence of malaria-helminth co-infection is high in children living in endemic countries. The nature of the interactions between malaria and helminth infection and the impact of the co-infection on anaemia remain inconclusive and may be modulated by the immune responses of the affected children.
Topics: Adolescent; Anemia; Animals; Child; Child, Preschool; Coinfection; Female; Helminthiasis; Helminths; Humans; Infant; Malaria; Male; Plasmodium; Prevalence; Soil
PubMed: 33600494
DOI: 10.1371/journal.pntd.0009138 -
Parasites & Vectors Feb 2021Ethiopia has set the ambitious national targets of eliminating soil-transmitted helminths (STH) and schistosomiasis (SCH) as public health problems by 2020, and breaking...
BACKGROUND
Ethiopia has set the ambitious national targets of eliminating soil-transmitted helminths (STH) and schistosomiasis (SCH) as public health problems by 2020, and breaking their transmission by 2025. This systematic review was performed to provide insight into the progress made by the national STH and SCH control programme purposed with reaching these targets.
METHODS
Studies published on STH and SCH in Ethiopia were searched for using Web of Science, PubMed, Scopus, and the resulting references of selected studies. Prevalence and intensity were analysed, stratified by region, age, and diagnostics.
RESULTS
A total of 231 papers published between 2000 and 2020 were included. Over the past two decades, Trichuris trichiura (TT) infection has shown the most statistically significant decrease (93%, p < 0.0001), followed by Schistosoma mansoni (SM) (69%, p < 0.0001), Ascaris lumbricoides (AL) (67%, p < 0.0001) and Schistosoma haematobium (83%, p = 0.038) infections. Geographically, parasite burden has only consistently shown a significant reduction in the Southern Nations, Nationalities and Peoples' Region of Ethiopia, where AL, TT, hookworm and SM significantly decreased by 80% (p = 0.006), 95% (p = 0.005), 98% (p = 0.009) and 87% (p = 0.031), respectively. Prevalence of STH was highest among adults across all species, contrary to typical age-infection profiles for TT and AL that peak among school-aged children. Expanding treatment to the whole community would target reservoirs of adult and preschool-aged infection within the community, assisting Ethiopia in reaching their national transmission break targets. There was substantial heterogeneity in diagnostic methods used across studies, the majority of which predominantly used single-slide Kato-Katz. This low slide frequency provides poor diagnostic sensitivity, particularly in low endemic settings.
CONCLUSION
The prevalence of STH and SCH in Ethiopia has decreased over time due to the strategic use of anthelmintics. Both standardising and increasing the sensitivity of the diagnostics used, alongside the ubiquitous use of parasite intensity with prevalence, would enable a more accurate and comparable understanding of Ethiopia's epidemiological progress. Further work is needed on community-wide surveillance in order to understand the burden and subsequent need for treatment among those outside of the standard school-based control program.
Topics: Cross-Sectional Studies; Ethiopia; Feces; Helminthiasis; Humans; Prevalence; Schistosomiasis; Soil; World Health Organization
PubMed: 33546757
DOI: 10.1186/s13071-021-04600-0 -
Tropical Diseases, Travel Medicine and... Feb 2021Schistosomiasis is a common helminthic infection in the tropics and subtropics, particularly in sub-Saharan African countries including Ethiopia. In these counties,... (Review)
Review
BACKGROUND
Schistosomiasis is a common helminthic infection in the tropics and subtropics, particularly in sub-Saharan African countries including Ethiopia. In these counties, Schistosoma mansoni infection is a significant public health problem due to the risk of reinfection and recurrent disease despite implementing several rounds preventive chemotherapy. This systematic review and meta-analysis aimed at assessing the pooled prevalence of schistosomiasis in Ethiopia.
METHODS
The PRISMA guidelines were followed to perform the systematic review and meta-analysis. Published studies from January 1999 to June 2020 were searched in Medline, PubMed, Google Scholar, EMBASE, HINARI, and Cochrane Library using key words including: "prevalence", "incidence", "schistosomiasis" "Bilharziasis", "Bilharzia", "S. mansoni ", "Ethiopia". Heterogeneity of included studies was assessed using Cochran's Q test and I test statistics while publication bias was assessed using Egger's test.
RESULTS
Ninety-four studies were included in the systematic review and meta-analysis. The pooled prevalence of S. mansoni in Ethiopia was 18.0% (95%CI: 14.0-23.0). The southern region of Ethiopia had a higher S. mansoni prevalence of 25.9% (995% CI, 14.9-41.1) than the national prevalence. The burden of S. mansoni infection was also higher than the national average in rural areas and among men with pooled prevalence of 20.2% (95% CI, 13.2-28.5) and 28.5% (95%CI, 22.7,35.1), respectively. The trend analysis showed that the prevalence of S. mansoni infection in Ethiopia decreased over the past 15 years, potentially because of the repeated preventive chemotherapy.
CONCLUSION
The review unveiled a moderate prevalence of S. mansoni infection in Ethiopia. Targeted treatment of at-risk population groups ad high burden areas coupled with implementation of integrated vector control strategies are critical to address the burden of Schistosomiasis.
PubMed: 33522949
DOI: 10.1186/s40794-020-00127-x -
PLoS Neglected Tropical Diseases Jan 2021Schistosomiasis affects nearly 220 million people worldwide, mainly in Sub-Saharan Africa (SSA). Preventive chemotherapy (PC) treatment, through regular mass-drug...
BACKGROUND
Schistosomiasis affects nearly 220 million people worldwide, mainly in Sub-Saharan Africa (SSA). Preventive chemotherapy (PC) treatment, through regular mass-drug administration (MDA) of Praziquantel tablets remains the control measure of choice by Ministries of Health. Current guidelines recommend that 75% of school-aged children receive treatment. Many programmes, however, struggle to achieve this target. Given the risk of high reinfection rates, attaining sustained high levels of treatment coverage is essential. This study provides a comprehensive review of the barriers and facilitators operating at different levels of analysis, from the individual to the policy level, conditioning the uptake of PC for schistosomiasis in SSA.
METHODOLOGY/PRINCIPAL FINDINGS
A systematic literature search was conducted in several databases for publications released between January 2002 and 2019 that examined factors conditioning the uptake of Praziquantel in the context of MDA campaigns in SSA. A total of 2,258 unique abstracts were identified, of which 65 were selected for full text review and 30 met all eligibility criteria. Joanna Briggs Institute's Critical Appraisal and the Mixed-Methods Assessment tools were used to assess the strength of the evidence. This review was registered with PROSPERO (CRD42017058525). A meta-synthesis approach was used. Results indicated publication bias, with the literature focusing on East African rural settings and evidence at the individual and programmatic levels. The main influencing factors identified included material wellbeing, drug properties, knowledge and attitudes towards schistosomiasis and MDAs, fears of side effects, gender values, community and health systems support, alongside programme design features, like training, sensitisation, and provision of incentives for drug-distributors. The effect of these factors on determining Praziquantel uptake were explored in detail.
CONCLUSIONS/SIGNIFICANCE
Multiple determinants of treatment uptake were found in each level of analysis examined. Some of them interact with each other, thus affecting outcomes directly and indirectly. The promotion of context-based transdisciplinary research on the complex dynamics of treatment uptake is not only desirable, but essential, to design effective strategies to attain high levels of treatment coverage.
Topics: Africa South of the Sahara; Animals; Anthelmintics; Chemoprevention; Health Knowledge, Attitudes, Practice; Humans; Mass Drug Administration; Praziquantel; Schistosoma; Schistosomiasis
PubMed: 33465076
DOI: 10.1371/journal.pntd.0009017 -
PloS One 2020Following the adoption of the World Health Assembly Resolution WHA 65.21 and Neglected Tropical Diseases road map 2021-2030, schistosomiasis control programmes have... (Meta-Analysis)
Meta-Analysis
INTRODUCTION
Following the adoption of the World Health Assembly Resolution WHA 65.21 and Neglected Tropical Diseases road map 2021-2030, schistosomiasis control programmes have shifted from morbidity control to disease elimination. However, several gaps continue to be observed in the implementation of control programmes with certain age groups omitted from these campaigns increasing health inequalities and risks of reinfections to previously treated groups. We used the Inverse Variance Heterogeneity (IVhet) model to estimate the prevalence of schistosomiasis infection among preschool-aged children.
METHODS
We did a systematic review of peer-reviewed literature on schistosomiasis in sub-Saharan Africa for the period January 1, 2000 to November 30, 2020. Quantitative data for cases of schistosomiasis infection were extracted, including country and region where the studies were done, year of publication and specific schistosome species observed. The IVhet model was used to estimate the pooled prevalence estimate (PPE), the heterogeneity and publication bias.
RESULTS
We screened 2601 articles to obtain 47 eligible studies containing quantitative data on preschool-aged children. Of the selected studies, 44.7% (n = 22) were from East Africa while the least number of studies obtained (2.1%, n = 1) was from Central Africa. 21712 subjects were screened for infection due to Schistosoma spp; 13924 for S. mansoni and 7788 for S. haematobium. The PPE for schistosomiasis among PreSAC was 19% (95% CI: 11-28). Infection due to S. mansoni (IVhet PPE: 22% (95% CI: 9-36) was higher than that due to S. haematobium (15%; 95% CI: 6-25). A Luis Furuya-Kanamori index of 1.83 indicated a lack of publication bias. High level of heterogeneity was observed (I2 > 90%) and this could not be reduced through subgroup analysis.
CONCLUSION
Schistosomiasis infection among pre-school aged children 6 years old and below is high. This indicates the importance of including this age group in treatment programmes to reduce infection prevalence and long-term morbidities associated with prolonged schistosome infection.
Topics: Africa South of the Sahara; Child; Child, Preschool; Female; Humans; Infant; Male; Prevalence; Schistosomiasis
PubMed: 33373405
DOI: 10.1371/journal.pone.0244695