Haematobium-infected Individuals Research Articles

Test-treat-track-test-treat (5T) approach for Schistosoma haematobium elimination on Pemba Island, Tanzania

BackgroundAfter decades of praziquantel mass drug administration (MDA), several countries approach schistosomiasis elimination. Continuing MDA in largely uninfected populations no longer seems justified. Alternative interventions to maintain the gains or accelerate interruption of transmission are needed. We report results, strengths, and shortcomings of novel test-treat-track-test-treat (5T) interventions in low Schistosoma haematobium prevalence areas on Pemba, Tanzania.MethodsSchool- and household-based surveys were conducted in 2021 and 2022 to monitor the S. haematobium and microhematuria prevalence and assess the impact of interventions. In 2021, 5T interventions were implemented in 15 low-prevalence areas and included: (i) testing schoolchildren in primary and Islamic schools for microhematuria as a proxy for S. haematobium, (ii) treating positive children, (iii) tracking them to their households and to water bodies they frequented, (iv) testing individuals at households and water bodies, and (v) treating positive individuals. Additionally, test-and-treat interventions were implemented in the 22 health facilities of the study area.ResultsThe S. haematobium prevalence in the school-based survey in 15 low-prevalence implementation units was 0.5% (7/1560) in 2021 and 0.4% (6/1645) in 2022. In the household-based survey, 0.5% (14/2975) and 0.7% (19/2920) of participants were infected with S. haematobium in 2021 and 2022, respectively. The microhematuria prevalence, excluding trace results, in the school-based survey was 1.4% (21/1560) in 2021 and 1.5% (24/1645) in 2022. In the household-based survey, it was 3.3% (98/2975) in 2021 and 5.4% (159/2920) in 2022. During the 5T interventions, the microhaematuria prevalence was 3.8% (140/3700) and 5.8% (34/594) in children in primary and Islamic schools, respectively, 17.1% (44/258) in household members, and 16.7% (10/60) in people at water bodies. In health facilities, 19.8% (70/354) of patients tested microhematuria-positive.ConclusionsThe targeted 5T interventions maintained the very low S. haematobium prevalence and proved straightforward and feasible to identify and treat many of the few S. haematobium-infected individuals. Future research will show whether 5T interventions can maintain gains in the longer-term and expedite elimination.Trial registrationISRCTN, ISCRCTN91431493. Registered 11 February 2020, https://www.isrctn.com/ISRCTN91431493.

Assessing the Effects of One-Time Praziquantel Treatment on Urogenital Lesions Detected by Ultrasound in Schistosoma haematobium-Infected Individuals in Chad.

The objective was to determine the impact of a single dose of praziquantel on urogenital lesions caused by Schistosoma haematobium. Ultrasound (US) was performed on three age groups of subjects with a positive test for hematuria, with the first examination performed in November 2017 and a follow-up visit 7 months later. None of the subjects had previously received treatment. The participants were categorized into three distinct age groups: group 1 = 1-15 years, group 2 = 15-30 years, and group 3 = ≥ 30 years. A total of 250 people from these three groups underwent screening: 99 in group 1, 90 in group 2, and 61 in group 3, among whom 131 (52.4%) had at least one detectable lesion of the urogenital tract on US. Follow-up US after 7 months was possible in 60%, 67%, and 77% of the respective groups (with lesions). The anomalies disappeared in 80% of individuals in group 1, 76% of those in group 2, and 65% in group 3. With the exception of calcifications, most visible anomalies had been resolved. The total number of anomalies is low. Severe obstructive uropathy was not detected. We can conclude that single treatment with praziquantel is able to cure visible anomalies, with the exception of calcifications. The low rate of anomalies, compared with levels in the literature, is speculated to be due to undetected death by obstructive uropathy caused by S. haematobium. This requires further investigation.

Schistosomiasis Induces Persistent DNA Methylation and Tuberculosis-Specific Immune Changes.

Epigenetic mechanisms, such as DNA methylation, determine immune cell phenotype. To understand the epigenetic alterations induced by helminth coinfections, we evaluated the longitudinal effect of ascariasis and schistosomiasis infection on CD4+ T cell DNA methylation and the downstream tuberculosis (TB)-specific and bacillus Calmette-Guérin-induced immune phenotype. All experiments were performed on human primary immune cells from a longitudinal cohort of recently TB-exposed children. Compared with age-matched uninfected controls, children with active Schistosoma haematobium and Ascaris lumbricoides infection had 751 differentially DNA-methylated genes, with 72% hypermethylated. Gene ontology pathway analysis identified inhibition of IFN-γ signaling, cellular proliferation, and the Th1 pathway. Targeted real-time quantitative PCR after methyl-specific endonuclease digestion confirmed DNA hypermethylation of the transcription factors BATF3, ID2, STAT5A, IRF5, PPARg, RUNX2, IRF4, and NFATC1 and cytokines or cytokine receptors IFNGR1, TNFS11, RELT (TNF receptor), IL12RB2, and IL12B (p < 0.001; Sidak-Bonferroni). Functional blockage of the IFN-γ signaling pathway was confirmed, with helminth-infected individuals having decreased upregulation of IFN-γ-inducible genes (Mann-Whitney p < 0.05). Hypomethylation of the IL-4 pathway and DNA hypermethylation of the Th1 pathway was confirmed by Ag-specific multidimensional flow cytometry demonstrating decreased TB-specific IFN-γ and TNF and increased IL-4 production by CD4+ T cells (Wilcoxon signed-rank p < 0.05). In S. haematobium-infected individuals, these DNA methylation and immune phenotypic changes persisted at least 6 mo after successful deworming. This work demonstrates that helminth infection induces DNA methylation and immune perturbations that inhibit TB-specific immune control and that the duration of these changes are helminth specific.

Organomegaly in Mali before and after praziquantel treatment. A possible association with Schistosoma haematobium

Continuous exposure to schistosome-infested water results in acute and chronic morbidity in all ages. We analysed occurence of organomegaly via ultrasonography and investigated a possible additive effect of dual-dose drug administration in 401 Schistosoma haematobium infected individuals from a highly endemic area in Mali. Mean intensity of infection at baseline (22.0 eggs per 10 ml) was reduced to 0.22 eggs per 10 ml 9 weeks after treatment (both treatments combined). Odds of persistent infection among those given dual-dose treatment was 41% of that in people given single dose (b = 0.41; p = 0.05; 95% CI 0.17–1.00), but after two years, 70.7% of the 157 participants, who completed the survey, were re-infected with no significant difference in prevalence and intensity of infection between treatment groups. Resolution of organomegaly occurred in all age groups after treatment. A novel association between Schistosoma haematobium infection and moderate portal vein enlargement was found in 35% (n: 55). Severe portal vein diameter enlargement was found in 3.2%. After two years, moderate hepatomegaly was present in 50.6%, moderate splenomegaly in 45.6% and moderate portal vein diameter enlargement in 19%. A subsequent dose of PZQ did not provide any additional long-term advantages.

Schistosoma haematobium infection and asymptomatic bacteriuria in young South African females

Schistosoma haematobium eggs can induce lesions in the urinary and genital tract epithelia, as eggs pass through or get trapped in the tissue. Local inflammatory reactions induced by S. haematobium eggs might affect the ability of bacteria to establish mucosal super-infection foci.S. haematobium infection and asymptomatic bacteriuria can both portray haematuria, proteinuria and leukocyturia. This shared set of proxy diagnostic markers could fuel routine misdiagnosis in S. haematobium endemic areas. Furthermore, S. haematobium infected individuals might be at a higher risk of contracting bacterial urinary tract infections, which could manifest either as symptomatic or asymptomatic bacteriuria.The aim of the current study was to explore whether schistosomal lesions are susceptible to super-infection by bacteria measured as asymptomatic bacteriuria. S. haematobium infection was determined by microscopy of urine samples. Furthermore, urine samples were tested with dipslides for asymptomatic bacteriuria and with dipsticks for haematuria, proteinuria and leukocytes. We found no association between asymptomatic bacteriuria and S. haematobium infection in a sample of 1040 female primary and high school students from a schistosomiasis endemic area in KwaZulu-Natal, South Africa. Furthermore, it was demonstrated that asymptomatic bacteriuria is not a bias for use of micro-haematuria as a proxy diagnostic measure for S. haematobium infection in this population.

Mannose-Binding Lectin and Susceptibility to Schistosomiasis

Background. Human ficolin 2 (encoded by FCN2) and mannose-binding lectin (encoded by MBL2) bind to specific pathogen-associated molecular patterns, activate the complement lectin cascade in a similar manner, and are associated with several infectious diseases. Our recently published study established certain FCN2 promoter variants and ficolin-2 serum levels as protective factors against schistosomiasis. Methods. We used the Nigerian cohort from our recently published study, which included 163 Schistosoma haematobium–infected individuals and 183 matched healthy subjects, and investigated whether MBL deficiency and MBL2 polymorphisms are associated with schistosomiasis. Results. MBL serum levels were significantly higher in controls and were associated with protection (P < .0001). The −550H minor allele was significantly associated with protection (P = .03), and the heterozygous genotypes −550HL were observed to confer protection (P = .03). The MBL2*HYPA haplotype was significantly associated with protection (P = .03), with significantly higher serum MBL levels in controls (P = .00073). The heterozygous 6-bp deletion in the promoter was observed to be a susceptibility factor in schistosomiasis (P = .03). Conclusions. In agreement with findings from our recently published study, the findings reported here support the observation that MBL is also associated with protection in schistosomiasis.

Ficolin-2 Levels and FCN2 Genetic Polymorphisms as a Susceptibility Factor in Schistosomiasis

Human ficolin-2 (L-ficolins) encoded by the FCN2 gene are pattern-recognition proteins involved in innate immunity and are associated with several infectious diseases. A Nigerian cohort of 168 Schistosoma haematobium-infected individuals and 192 healthy controls were examined for functional single-nucleotide polymorphisms in the promoter region (-986G>A, -602G>A, -4A>G) and in exon 8 (+6424G>T) using real-time polymerase chain reaction. The FCN2 -986A and -4G alleles were significantly associated with the occurrence of schistosomiasis (P = .0004 for -986G>A; P = .0001 for -4A>G). The heterozygous genotypes (P = .0006 for -986G>A; P = .0002 for -4A>G) were observed to be a risk factor for susceptibility to schistosomiasis, whereas the homozygous genotypes of major alleles (P = .0002 for -986G>A; P = .0001 for -4A>G) were observed to shield against schistosomiasis. The haplotype AGGG (P = .0002) was observed to be a risk factor for susceptibility to schistosomiasis compared with controls, and the haplotype GGAG (P = .04) was observed to confer protection compared with patients. Ficolin-2 serum level was significantly higher in controls (P < .005) and in controls with GGAG haplotypes (P < .0001). Our findings demonstrate that FCN2 promoter variants (-986G>A and -4A>G) influence ficolin-2 serum levels and susceptibility to schistosomiasis.

Novel Automated Biomarker Discovery Work Flow for Urinary Peptidomics

Urine is potentially a rich source of peptide biomarkers, but reproducible, high-throughput peptidomic analysis is often hampered by the inherent variability in factors such as pH and salt concentration. Our goal was to develop a generally applicable, rapid, and robust method for screening large numbers of urine samples, resulting in a broad spectrum of native peptides, as a tool to be used for biomarker discovery. Peptide samples were trapped, desalted, pH-normalized, and fractionated on a miniaturized automatic reverse-phase strong cation exchange (RP-SCX) cartridge system. We analyzed eluted peptides using MALDI-TOF, Fourier transform ion cyclotron resonance, and liquid chromatography-iontrap mass spectrometry. We determined qualitative and quantitative reproducibility of the system and robustness of the method using BSA digests and urine samples, and we used a selected set of urine samples from Schistosoma haematobium-infected individuals to evaluate clinical applicability. The automated RP-SCX sample cleanup and fractionation system exhibits a high qualitative and quantitative reproducibility, with both BSA standards and urine samples. Because of the relatively high cartridge binding capacity (1-2 mL urine), eluted peptides can be measured with high sensitivity using multiple mass spectrometric techniques. As proof of principle, hemoglobin-derived peptides were identified in urine samples from S. haematobium-infected individuals, even when the microhematuria test was negative. We present a practical, step-by-step method for screening and identification of urinary peptides. Alongside the analytical method evaluation on standard samples, we demonstrate its feasibility with actual clinical material.

Use of monoclonal antibodies prepared against Schistosoma mansoni hatching fluid antigens for demonstration of Schistosoma haematobium circulating egg antigens in urine.

A panel of 17 monoclonal antibodies (MAbs) against Schistosoma soluble egg antigens (SEAs) was produced from BALB/c mice immunized with antigens secreted/excreted by Schistosoma mansoni eggs. In this study, we demonstrate that 16 MAbs were reactive with S. haematobium SEA in addition to S. mansoni SEA. The MAbs were tested as potential immunodiagnostic reagents in a homologous sandwich ELISA format to detect circulating soluble egg antigens (CSEAs) in serum and urine samples of S. mansoni- or S. haematobium-infected individuals. When samples of S. mansoni-infected individuals were tested, none of these MAbs performed as good as the previously described S. mansoni-specific 114-5B1-A and 114-4D12-A MAbs. However, 11 MAbs (of the IgM isotype) detected CSEA in urine samples of S. haematobium-infected individuals. Three MAbs, 290-2E6-A, 291-3D5-A, and 291-5D5-A, were selected for a pilot study with 47 urine samples of S. haematobium-infected individuals from Kenya. The CSEA levels detected with each of these ELISAs showed a significant correlation with urinary egg counts (Spearman rho > 0.37, P < 0.01) and with each other (Spearman rho > 0.74, P < 0.001). Based on the 92% specificity and 90% sensitivity of the assay, the ELISA using MAb 290-2E6-A was found to be the most promising assay for immunodiagnosis of S. haematobium infections.

Elevated proliferation and interleukin-4 release from CD4+ cells after chemotherapy in human Schistosoma haematobium infection.

Cellular immune responses to schistosomal antigens were examined in 110 Schistosoma haematobium-infected individuals before and 5 weeks after treatment with praziquantel. Chemotherapy resulted in significant decrease in worm load as measured by egg output and circulating antigens. The proliferative responses to adult worm antigen (AWA) increased significantly after treatment (p < 0.001) whereas purified protein derivative of tuberculin or phytohemagglutinin responsiveness was unaffected. Interleukin (IL)-4 production in response to both AWA and soluble egg antigen (SEA) increased markedly after treatment (p < 0.001), but IL-5 remained unchanged. None of the studied subjects released any measurable IL-2 and only 21% produced interferon (IFN)-gamma in response to parasite antigens. The deficiency in either IFN-gamma or IL-2 release was not restored by chemotherapy. Thus chronic infection with S. haematobium is associated with the reversible down-regulation of T cell proliferative responses and IL-4 release. Results from cell depletion experiments indicated that CD4+ T cells were the target of this down-modulation.

Association of bacteriuria and urinary nitrosamine formation with Schistosoma haematobium infection in the Qalyub area of Egypt

In Egypt, bladder cancer incidence is high in areas where the prevalence and intensity of Schistosoma haematobium infection is also high. Experimental evidence shows bladder carcinogenesis to be a multi-stage process which can be accelerated by many factors. N-nitroso compounds, some of which are known bladder carcinogens, can be formed from amine precursors and nitrate in urine during some bacterial infections. In experimental animals the growth of nitrosamine-induced urothelial cancers is accelerated by damage to the urothelium caused by S. haematobium infections, and by analogy in man this could account for the lower peak age of incidence of this cancer in Egypt by comparison with Europe. The present study was designed to investigate whether bacterial infection of the urinary tract was common in areas of endemic schistosomiasis and whether N-nitrosamines were regularly found to be associated with bacteriuria. Urine samples from young men in the Qalyub area of Egypt and from an adjacent Delta region were analysed for S. haematobium ova, the nature and intensity of any bacterial infection, nitrate and nitrite, and total N-nitroso compounds plus volatile N-nitrosamines. A relatively high prevalence of bacteriuria was found in young men with schistosomiasis and low levels of N-nitroso compounds were present in all specimens. When the groups were sub-divided on the basis of the ability of their bacterial flora to reduce nitrate to nitrite (the latter is required for the nitrosation of amine precursors to N-nitroso compounds), significantly higher levels of N-nitroso compounds were found in S. haematobium-infected individuals also infected with nitrate-reducing bacteria by comparison either with uninfected controls (p < 0 · 0005) or with those infected with non-nitrate-reducing bacteria (p <0 · 001). The results show N-nitroso compounds to be present in the urines of young men in areas of endemic S. haematobium infection in Egypt, and elevated levels of urinary N-nitroso compounds to be associated with infection of the urinary tract by various species of nitrate-reducing bacteria.

The circumoval precipitin test for the serodiagnosis of human schistosomiasis mansoni and haematobia.

The circumoval precipitin (COP) test was used to detect serum antibodies to Schistosoma mansoni, S. haematobium, or both species by using eggs of either species of schistosome. Eggs of either species were adequate as antigens but, in general, the sera of S. mansoni-infected individuals caused more circumoval precipitates for form around S. mansoni eggs (higher reactivity) than did the sera of S. haematobium-infected individuals. However, sera from individuals with either or double infections reacted equally with S. haematobium eggs. In addition, more circumoval precipitates were formed around S. haematobium eggs obtained from human urine than around S. mansoni eggs obtained from mouse livers. We demonstrate that human infection with either species of schistosome can be diagnosed by the COP test, but that identification of the schistosome species involved in the infection is not possible with this test. In addition, by eliminating the need for maintaining a schistosome life cycle in the laboratory the demonstration of the high reactivity of S. haematobium eggs from urine greatly simplifes the performance of the COP test in areas where S. haematobium-infected individuals can be found and will cooperate by providing urine. We also obtained serum specimens by venipuncture, and capillary blood dried on filter paper, from Egyptians infected with S. mansoni, S. haematobium, or with both species of schistosome. Serum eluted from the filter paper was then compared with serum from venipunctures with respect to their reactivity in the COP test done with fresh S. haematobium eggs as antigen. Serum eluates from freshly dried blood were significantly less reactive than serum obtained by venipuncture in terms of positive reactions and characteristics of the COP precipitate. Storage of the filter paper blood at 4°C for 6–12 months resulted in an even greater reduction of reactivity. Thus, for the COP test serum obtained by venipuncture is preferable to serum eluates obtained from blood on filter paper for the serodiagnosis of infection with S. mansoni and S. haematobium.