Elimination of schistosomiasis requires multifactorial diagnostics: evidence from high- and low-prevalence areas in the Nile Delta, Egypt.

BACKGROUND: Schistosomiasis is one of the neglected tropical diseases (NTDs) selected for worldwide elimination in the near future. Egypt has made strong progress against its two endemic species of Schistosoma mansoni and S. haematobium. The former is prevalent in the Nile Delta with the latter dominating in the Nile south of Cairo. Innovative efforts are needed to reach the goal as further reduction of the prevalence has stalled due to ongoing transmission. In this study we aimed to explore the difference between low and high prevalence villages with regard to knowledge attitude and practice about schistosomiasis, utilization of health services, infection and transmission indices. METHODS: A hybrid cross-sectional longitudinal study was conducted with three annual follow-ups conducted during 1994-1996. We used a representative systematic random sampling technique investigating 993 individuals from the high prevalence village and 614 from the low prevalence village. Data were analyzed using SPSS, comparing proportions with the Chi square test and means with the Student t test, and ANOVA. RESULTS: Compliance of faecal sampling and chemotherapy was above 70% in both villages over the whole study period. Selective praziquantel treatment resulted in a significant reduction of prevalence and intensity of infection in both villages, dropping from 35.8% prevalence to 20.6%, in the low-prevalence village, and from 69.5 to 45.9% in the high-prevalence one. Intensity of infection at the base line was 30 eggs per gram (EPG) of stool in the low-prevalence village versus 105 EPG in the high-prevalence village. However, after the second round, reinfection rebounded by 22% in the high-prevalence village, while a slight improvement of the infection indices was demonstrated in the low-prevalence one. The level of knowledge was modest in both villages: people knew about self-protection and treatment, but not much about the role of human excreta for schistosomiasis transmission. While all participants maintained that using the water from the canals was inevitable, inhabitants in the high-prevalence village showed significantly lower scores reflecting higher water contact compared to the low-prevalence one. Many of them (67%) did not utilize the health centre at all compared to 26% of the people in the low-prevalence village. Interestingly, private clinics were seen as the primary source of health care by both villages, but more frequently so in the high-prevalence village (used by 87.2% of the inhabitants) compared to the low-prevalence one (59.8%). CONCLUSIONS: Even if chemotherapy works well as reflected by the observed downregulation of intensity of infection in both villages, reinfection continued due to difficulties to avoid water contact. Efforts must be made to make people understand the role of human excreta for transmission. There is also a need to make people better trust the medical services available.

Prophylactic effect of artemether on human schistosomiasis mansoni among Egyptian children: A randomized controlled trial.

A double-blind, randomized controlled trial was conducted in an endemic focus for Schistosoma mansoni in Kafr El-Sheikh Governorate, Northern Nile Delta, Egypt, to evaluate the prophylactic effect of artemether (ART) given in conjunction with praziquantel (PZQ). The study encompassed 913 primary school children randomly assigned to two treatment groups PZQ/ART and PZQ/ART-placebo. At baseline, both groups received 40 mg/kg body weight of PZQ twice four weeks apart, after which one group received 6 mg/kg body weight of ART every 3 weeks in 5 cycles during the transmission season and the other group received ART-placebo. At the end of the study, prevalence of infection among the PZQ/ART was approximately half that of the PZQ/ART-placebo group, i.e. 6.7% versus 11.6%, and incidence of new infections for the PZQ/ART was 2.7% versus 6.5% for the PZQ/ART-placebo. In conclusion, PZQ/ART combined therapy might be considered as an adjunct measure against human schistosomiasis, by specifically reducing transmission and therefore contribute to disease elimination.

Can human schistosomiasis mansoni control be sustained in high-risk transmission foci in Egypt?

BACKGROUND: Control of human schistosomiasis remains a longstanding issue on the agenda of the Egyptian Ministry of Health and Population (MOHP). Substantial impact on morbidity and prevalence of S. mansoni was widely reported after the National Schistosomiasis Control Program (NSCP) extended selective treatment with praziquantel (PZQ) to the Nile Delta in 1992 and upgrading this approach to mass drug administration (MDA) in 1997. Disease elimination, however, eludes NSCP as the micro-level includes many high-risk foci that sustain transmission, which has not been subjected to investigation. METHODS: The study included five high-risk Nile Delta villages situated in the Kafr El-Sheikh Governorate. The total sample size amounted to 2382 individuals of both sexes and all ages. Diagnosis was based on four Kato-Katz slides from two consecutive stool samples. Data were investigated using SPSS, comparing proportions with the Chi square test and means with the Student t test, while strength of the associations were subjected to Odds Ratio (OR) analysis. RESULTS: The overall prevalence of schistosomiasis in the study area was found to be 29%, while the mean geometric mean egg count (GMEC) was low (66.78 ± 4.4) indicating low intensity of infection. The mean village prevalence rates ranged from 16.5% to 49.5% and the GMECs from 35.2 to 86.2 eggs per gram (EPG) of stool. The difference of prevalence between villages was statistically significant at P < 0.05, and the prevalence was significantly higher among males than among females, P < 0.05, OR =1.4 and 95% CI (1.16-1.60). Infection peaked in the next youngest age group (5- ≤ 10 years of age) at an average prevalence of 50.8% with the GMEC reaching 209 EPG of stool in the village with the highest prevalence. The average prevalence and GMEC among children <5 years were 20.6% and 92.7 EPG, respectively. CONCLUSION: Transmission of S mansoni in high-risk areas in the Nile Delta remains uninterrupted calling for improved, more comprehensive control strategies. Further investigations are needed to find out whether these results are due to inefficacy of PZQ, surviving immature worms or drug resistance.

Efficacy and safety of arachidonic acid for treatment of school-age children in Schistosoma mansoni high-endemicity regions.

Arachidonic acid (ARA), an omega-6 fatty acid, is a potent schistosomicide that displayed significant and safe therapeutic effects in Schistosoma mansoni-infected schoolchildren in S. mansoni low-prevalence regions. We here report on ARA efficacy and safety in treatment of schoolchildren in S. mansoni high-endemicity areas of Kafr El Sheikh, Egypt. The study was registered with ClinicalTrials.gov (NCT02144389). In total, 268 schoolchildren with light, moderate, or heavy S. mansoni infection were assigned to three study arms of 87, 91, and 90 children and received a single dose of 40 mg/kg praziquantel (PZQ), ARA (10 mg/kg per day for 15 days), or PZQ combined with ARA, respectively. The children were examined before and after treatment for stool parasite egg counts and blood biochemical, hematological, and immunological parameters. ARA, like PZQ, induced moderate cure rates (50% and 60%, respectively) in schoolchildren with light infection and modest cure rates (21% and 20%, respectively) in schoolchildren with high infection. PZQ and ARA combined elicited 83% and 78% cure rates in children with light and heavy infection, respectively. Biochemical and immunological profiles were either unchanged or ameliorated after ARA therapy. Combination of PZQ and ARA might be useful for treatment of children with schistosomiasis in high-endemicity regions.

Efficacy and safety of arachidonic acid for treatment of Schistosoma mansoni-infected children in Menoufiya, Egypt.

Arachidonic acid (ARA), an omega-6 fatty acid, kills juvenile and adult schistosomes in vitro and displays highly significant and safe therapeutic effects in mice and hamsters infected with Schistosoma mansoni or S. haematobium. This study aims to examine the efficacy and safety of ARA in treatment of school-age children infected with S. mansoni. In total, 66 S. mansoni-infected schoolchildren (20-23 children/study arm) received a single dose of 40 mg/kg praziquantel (PZQ), ARA (10 mg/kg per day for 15 days), or PZQ combined with ARA. The children were examined before and after treatment for worm egg counts in stool and blood biochemical and immunological parameters. ARA proved to be as efficacious as PZQ in treatment of schoolchildren with low infection intensity (78% and 85% cure rates, respectively). For moderate-intensity infection, the ARA and PZQ combination led to 100% cure rate. Biochemical, hematological, and immunological parameters were either unchanged or ameliorated after ARA therapy.

Epidemiology of Schistosomiasis in Egypt: Travel through Time: Review.

Schistosomiasis is a parasitic disease caused by blood flukes (Trematodes) of the genus Schistosoma (S.). It is well documented that schistosomiasis haematobium was endemic in Ancient Egypt. Infection was diagnosed in mummies 3000, 4000 and 5000 years old. Scott was the first to describe the pattern of schistosomiasis infection in Egypt. Schistosomiasis haematobium was highly prevalent (60%) both in the Nile Delta and Nile Valley South of Cairo in districts of perennial irrigation while it was low (6%) in districts of basin irrigation. Schistosoma mansoni infected 60% of the population in the Northern and Eastern parts of the Nile Delta and only 6% in the Southern part. Neither S. mansoni cases nor its snail intermediate host were found in the Nile Valley South of Cairo. The building of the Aswan High Dam -which was completed in 1967 - did not cause any increase in schistosomiasis prevalence. In 1990, a study conducted in nine governorates of Egypt confirmed the change in the pattern of schistosomiasis transmission in the Delta. There was an overall reduction in S. mansoni prevalence while Schistosoma haematobium had continued to disappear. In Middle and Upper Egypt there was consistent reduction in the prevalence of S. haematobium except in Sohag, Qena, and Aswan governorates. However, foci of S. mansoni were detected in Giza, Fayoum, Menya and Assiut. All schistosomiasis control projects implemented in Egypt from 1953 to 1985 adopted the strategy of transmission control and were based mainly on snail control supplemented by anti-bilharzial chemotherapy. In 1997, the National Schistosomiasis Control Program (NSCP) was launched in the Nile Delta. It adopted morbidity control strategy with Praziquantel mass treatment as the main component. In 1996, before the NSCP, 168 villages had S. mansoni prevalence >30%, 324 villages 20-30% and 654 villages 10-20%. By the end of 2010, in the whole country only 29 villages had prevalence >3% and none had more than 10%.

A research agenda for helminth diseases of humans: diagnostics for control and elimination programmes.

Diagnostic tools appropriate for undertaking interventions to control helminth infections are key to their success. Many diagnostic tests for helminth infection have unsatisfactory performance characteristics and are not well suited for use in the parasite control programmes that are being increasingly implemented. Although the application of modern laboratory research techniques to improve diagnostics for helminth infection has resulted in some technical advances, uptake has not been uniform. Frequently, pilot or proof of concept studies of promising diagnostic technologies have not been followed by much needed product development, and in many settings diagnosis continues to rely on insensitive and unsatisfactory parasitological or serodiagnostic techniques. In contrast, PCR-based xenomonitoring of arthropod vectors, and use of parasite recombinant proteins as reagents for serodiagnostic tests, have resulted in critical advances in the control of specific helminth parasites. The Disease Reference Group on Helminths Infections (DRG4), established in 2009 by the Special Programme for Research and Training in Tropical Diseases (TDR) was given the mandate to review helminthiases research and identify research priorities and gaps. In this review, the diagnostic technologies relevant to control of helminth infections, either available or in development, are reviewed. Critical gaps are identified and opportunities to improve needed technologies are discussed.

A research agenda for helminth diseases of humans: towards control and elimination.

Human helminthiases are of considerable public health importance in sub-Saharan Africa, Asia, and Latin America. The acknowledgement of the disease burden due to helminth infections, the availability of donated or affordable drugs that are mostly safe and moderately efficacious, and the implementation of viable mass drug administration (MDA) interventions have prompted the establishment of various large-scale control and elimination programmes. These programmes have benefited from improved epidemiological mapping of the infections, better understanding of the scope and limitations of currently available diagnostics and of the relationship between infection and morbidity, feasibility of community-directed or school-based interventions, and advances in the design of monitoring and evaluation (M&E) protocols. Considerable success has been achieved in reducing morbidity or suppressing transmission in a number of settings, whilst challenges remain in many others. Some of the obstacles include the lack of diagnostic tools appropriate to the changing requirements of ongoing interventions and elimination settings; the reliance on a handful of drugs about which not enough is known regarding modes of action, modes of resistance, and optimal dosage singly or in combination; the difficulties in sustaining adequate coverage and compliance in prolonged and/or integrated programmes; an incomplete understanding of the social, behavioural, and environmental determinants of infection; and last, but not least, very little investment in research and development (R&D). The Disease Reference Group on Helminth Infections (DRG4), established in 2009 by the Special Programme for Research and Training in Tropical Diseases (TDR), was given the mandate to undertake a comprehensive review of recent advances in helminthiases research, identify research gaps, and rank priorities for an R&D agenda for the control and elimination of these infections. This review presents the processes undertaken to identify and rank ten top research priorities; discusses the implications of realising these priorities in terms of their potential for improving global health and achieving the Millennium Development Goals (MDGs); outlines salient research funding needs; and introduces the series of reviews that follow in this PLoS Neglected Tropical Diseases collection, "A Research Agenda for Helminth Diseases of Humans."

Efficacy of two praziquantel treatments among primary school children in an area of high Schistosoma mansoni endemicity, Nile Delta, Egypt.

Praziquantel is the cornerstone of schistosomiasis control. A number of reports from endemic areas suggest that resistance or tolerance to praziquantel might exist in Schistosoma mansoni. Several explanations were postulated. The present work was designed to test the hypothesis that a low praziquantel (pzq) cure rate in Egypt is due to survival and maturation of immature stages that escaped pzq, which is effective against mature S. mansoni worms only. The study sample included 1351 children attending El Rouse primary school located in El Rouse village, Nile Delta, Egypt. All children received 2 pzq doses (40 mg/kg) 4 weeks apart. Diagnosis of S. mansoni infection and cure assessment were based on examination of 2 Kato slides prepared from a single stool sample collected before and 4 weeks after the first and second treatments. The cure rate was 78·8% after the first treatment and increased significantly to 90·8% after the second treatment. Egg reduction rates were 71·2% and 77·2% after 1 and 2 treatments respectively. Pre-treatment intensity of infection has a great influence on cure and egg reduction rates. Our results confirmed that low praziquantel cure rate, in Egypt, might be attributed, even partially, to survival and maturation of the immature S. mansoni stages that escaped pzq that is effective against mature worms only.

Triclabendazole and its two main metabolites lack activity against Schistosoma mansoni in the mouse model.

Some have claimed that triclabendazole, a safe and efficacious drug for the treatment of fascioliasis, also exhibits antischistosomal properties, but results are conflicting. We assessed the effect of triclabendazole and its two main metabolites against two different strains of Schistosoma mansoni harbored in mice. Low worm burden reductions (18.6-35.9%) were observed in mice infected with an Egyptian strain of S. mansoni and treated with a single dose of 120 mg/kg 3 days before infection or single/double doses of 120-200 mg/kg 7 weeks after infection. Triclabendazole failed to significantly reduce hepatic and intestinal tissue egg loads, and eggs of all developmental stages were observed. Administration of 400 mg/kg of either triclabendazole, triclabendazole sulphone, or triclabendazole sulfphoxide to mice infected with a Liberian strain of S. mansoni resulted in worm burden reductions < 10%. In comparison, high worm burden reductions (82-100%) were observed in S. mansoni-infected mice treated with single oral doses of 400, 500, or 500 mg/kg twice a day praziquantel, regardless of the S. mansoni strain. We conclude that triclabendazole and its main metabolites display weak and inconsistent schistosomicidal activities.

Efficacy of myrrh in the treatment of human Schistosomiasis mansoni.

Myrrh (Mirazid) has been produced and marketed as an antischistosomal drug since 2001. The current study was designed to assess the efficacy of a commercially available product of myrrh. One hundred four individuals, infected with Schistosoma mansoni, were randomized in two groups, one for myrrh and the second for praziquantel. Treatment-whether myrrh or praziquantel-was given twice with a 3-week interval. The cure rate with myrrh was very low, 15.6% after the first treatment, and 8.9% after the second treatment. Egg reduction among uncured persons was also very low, being 17.2% after the first treatment, and 28% after the second treatment. Praziquantel cure rate was 73.7% and 76.3%, and individuals still passing S. mansoni ova after praziquantel treatment showed a substantial reduction in the geometric mean egg counts (84% and 88.2% after the first and second treatments, respectively). When 34 individuals-uncured after two myrrh treatments-were offered praziquantel in the standard dose, 32 of them stopped passing S. mansoni eggs when tested 4 weeks post-treatment. The results of the current study raise serious doubts about the antischistosomal properties of Mirazid.

In vitro cellular and humoral responses to Schistosoma mansoni vaccine candidate antigens.

Few studies comparing schistosomiasis vaccine candidate antigens between laboratories have been carried out and published. Generally, only the investigators who discovered the molecules have evaluated them in either experimental animal models or in human correlate studies. In an attempt to identify responses against specific antigens and investigate their association with resistance versus susceptibility to re-infection, we studied the serological reactions and the cytokine responses stimulated by a panel of 10 candidate vaccine molecules in 225 long-term residents of an area endemic for Schistosoma mansoni in Egypt. The panel consisted of four recombinant antigens (Sm62-Irv5, Sm37-G3PDH, Sm28-GST and Sm14-FABP), one full-length native protein (Sm97-paramyosin), two synthetic peptides (MAP3 and MAP4) and three unpublished antigens (PR52-filamin, PL45-phosphoglycerate kinase, PN18-cyclophilin). Two different study designs, one based on retrospective and the other on prospective parasitological data were applied in the evaluation of the immune responses. Using historical data collected over the previous 5 years, correlations between frequency of re-infection and antigen-specific immune responses were investigated. In the prospective arm of the study, the subjects were followed over time after treatment with praziquantel with periodic immunological tests and stool examinations. Thus, highly specific humoral and cellular immune reactions in response to the 10 antigens described above could be correlated, both prospectively and retrospectively, with detailed epidemiological data covering a 66-month period. The immune response profiles produced were unique to each antigen but no clear "winner" or "winners" were identified. However, markers for both resistance and susceptibility to re-infection were identified for each molecule indicating which types of responses to aim for in vaccination and which ones to avoid. The insights gained from this approach should be useful for antigen selection and ultimately for vaccine formulation prior to Phase I/II trials in humans.

Blueprint for schistosomiasis vaccine development.

A number of different schistosome antigens are capable of partially protecting experimental animals from challenge infection. More than 100 such antigens have been identified, about 15% of which are strongly protective and deemed promising though they do not reach the level close to sterile immunity seen after vaccination with irradiated cercariae. Studies of human correlate reactions, i.e. serological reactions and cytokine responses to schistosomiasis antigens, in individuals living in areas endemic for schistosomiasis have shown associations between certain antigen-specific immune responses and lack of re-infection over time. This approach was applied in Brazil and Egypt where it was possible to epidemiologically follow cohorts of individuals in endemic areas for extended periods of time correlating infection status with immune responses against a panel of well-researched, highly purified vaccine candidates. The immune correlates found were unique to each antigen and could be either positive or negative, i.e. associated with resistance or with susceptibility to re-infection. However, few antigens were clear-cut in this respect, i.e. the majority of them induced ambiguous responses. For example, a single antigen might have a significant positive correlation when antigen-driven interferon (INF)-gamma production is measured but also show a significant negative correlation with respect to the IgG1 titre induced. These observations suggest that there are desirable, antigen-specific immune responses that would be valuable in a vaccine but they also indicate that there are responses that must be avoided. The insights gained should be useful not only for antigen selection but also for vaccine formulation prior to Phase I/II trials in humans. It would be of great value if similar independent, long-term human correlate studies could also be undertaken in areas endemic for Schistosoma japonicum.