[Schistosomiasis in areas of low endemicity: an overly neglected disease. The example of Schistosoma monsoni in BrazilEsquistossomose em áreas de baixa endemicidade, uma doença muito negligenciada. O exemplo de Schistosoma mansoni no Brasil]. / La schistosomose dans les zones de faible endémicité, une maladie trop négligée. Le cas de Schistosoma monsoni au Brésil.

Human schistosomiasis is a parasitic disease caused by an infection with trematodes of the genus Schistosoma. The disease mainly affects impoverished populations. Around 800 million people are exposed to the infection, which is a public health problem in the tropical and subtropical regions of Africa, Asia, the Caribbean and South America. In Brazil, Schistosoma mansoni is the only species that causes schistosomiasis and the disease is widely distributed. Conventional diagnosis of the disease is carried out by detecting eggs using parasitological methods, such as the Kato-Katz test. Schistosomiasis has been reported in all regions of Brazil and is characterized as endemic in seven states in the Northeast Region and two states in the Southeast Region. In 2015, 78,7% of all cases reported in Brazil occurred in the Northeast Region. It is estimated that 1,5 million people is infected with this disease in Brazil and more than 25 millions live in areas with a high risk of transmission. Despite the reduction in mortality and morbidity, schistosomiasis was responsible for 8,756 deaths between 2000 and 2011 and 2,517 deaths between 2015 and 2019 in Brazil and it remains an important public health problem. In the state of Rio de Janeiro, some areas have low endemicity or isolated foci of Schistosoma mansoni and the majority of infected individuals have mild infections. The last survey of the disease in the state of Rio de Janeiro was carried out between 2010 and 2015 in students aged 7 to 17.Schistosomiasis was reported in 10 of the 21 municipalities studied. Of the 5,111 school children screened for S. mansoni infection, 46 (1,65%) were tested positive. Studies carried out in areas of low endemicity in Rio de Janeiro showed that among the 205 patients infected by S. mansoni in Sumidouro, around 84% were aged 14 or over and all, except one individual, had the intestinal form (91,2%) or hepato-intestinal (8,3%) of schistosomiasis. Another study carried out in Sumidouro showed that with tests based on patent Schistosoma egg infection determined by the Kato-Katz test, active infections were diagnosed in eight (8/108) individuals. The intensity of infection expressed by parasite loads ranged from 6 to 72 eggs per gram of feces/individual. The results showed DNA amplification in 32 of the 100 individuals tested by real-time PCR. All individuals with patent ovo infection showed positive DNA amplification. These studies showed that if we only analyzed school-age children using the Kato-Katz test, the majority of the infected population would never be diagnosed with S. mansoni infection. In situations of low endemicity, with low intensities of infection, with low severity in the population and in the most affected age groups, schistosomiasis requires a more sensitive diagnostic approach (e.g. screening by PCR rather than Kato test), otherwise many infected individuals will remain invisible to the healthcare system.

A esquistossomose humana é uma doença parasitária causada por uma infecçâo por vermes sanguíneos do gènero Schistosoma. A doença afeta principalmente populaçoes empobrecidas. Cerca de 800 milhoes de pessoas estâo expostas à infecçâo, sendo um problema de saúde pública nas regioes tropicais e subtropicais de África, Ásia, Caribe e América do Sul. No Brasil, o Schistosoma mansoni é a única espécie causadora da esquistossomose e a doença é amplamente distribuida. O diagnóstico convencional da doença é realizado pela detecçâo dos ovos através de métodos parasitológicos, como o teste de Kato-Katz. A esquistossomose foi notificada em todas as regioes do Brasil, e é caracterizada como endèmica em sete estados da Regiâo Nordeste e dois estados da Regiâo Sudeste. Em 2015, 78,7% de todos os casos notificados no Brasil ocorreram na Regiâo Nordeste. Estima-se que 1,5 milhâo de pessoas estejam infectadas com esta doença no Brasil e mais de 25 milhoes vivam em áreas com alto risco de transmissâo. Apesar da reduçâo da mortalidade e morbidade, a esquistossomose foi relatada em 8.756 mortes entre 2000 e 2011 e em 2.517 mortes entre 2015 e 2019 no Brasil e continua sendo um importante problema de saúde pública. No Estado do Rio de Janeiro, algumas áreas apresentam baixa endemicidade ou focos isolados de Schistosoma mansoni e a maioria dos individuos infectados apresenta infecçoes leves. O último levantamento da doença no Estado do Rio de Janeiro foi realizado entre 2010 e 2015 em estudantes de 7 a 17 anos. A esquistossomose foi relatada em 10 dos 21 municipios estudados. Das 5.111 crianças escolares triadas para infecçâo por S. mansoni, 46 (1,65%) testaram positivo. Estudos realizados em áreas de baixa endemicidade no Rio de Janeiro mostraram que dentre os 205 pacientes infectados por S. mansoni em Sumidouro, cerca de 84% tinham 14 anos ou mais e todos, exceto um individuo, tinham a forma intestinal (91,2%) ou hepato-intestinal (8,3%) da esquistossomose. Outro estudo realizado em Sumidouro, mostrou que testes baseados em infecçâo patente de ovo de Schistosoma determinada pelo teste de Kato-Katz, infecçoes ativas foram diagnosticadas em oito (8/108) individuos. A intensidade de infecçâo expressa pelas cargas parasitárias variou de 6 a 72 ovos por grama de fezes/individuo. Os resultados mostraram amplificaçâo do DNA em 32 dos 100 individuos testados por PCR em tempo real. Todos os indivíduos com infecçâo ovo-patente apresentaram amplificaçâo de DNA positiva. Tais estudos mostraram que se analisarmos apenas crianças em idade escolar pelo teste de Kato-Katz, a maioria da populaçâo infectada nunca seria diagnosticada com infecçâo pelo S. mansoni. Em situaçoes de baixa endemicidade, com baixas intensidades de infecçâo, com baixa gravidade na populaçâo e nas faixas etárias mais afetadas, a esquistossomose requer uma abordagem diagnóstica mais sensivel (por exemplo, triagem por PCR em vez do teste de Kato), caso contràrio, muitos individuos infectados permanecerâo invisiveis para o sistema de saúde.

Descriptive study on risk of increased morbidity of schistosomiasis and graft loss after liver transplantation.

Solid-organ transplantation procedures have witnessed a surge in frequency. Consequently, increased attention to associated infections and their impact on graft success is warranted. The liver is the principal target for infection by the flatworm Schistosoma mansoni. Hence, rigorous screening protocols for this parasite should be implemented for liver transplantation donors and recipients. This study investigated the risks posed by schistosomiasis-infected liver tissues for successful liver transplantation (LT), considering donors and recipients, by analyzing reported cases. Among the 43 patients undergoing LT (donors = 19; recipients = 24), 32 were infected with S. mansoni, five were infected with other Schistosoma species, and no identification was made in four patients. Reported follow-up periods ranged from 1 to 132 months, and all patients achieved successful recovery. As these helminths do not replicate in their vertebrate hosts, immunosuppressive treatment is not expected to promote increased morbidity or reactivation. Moreover, suspected or confirmed schistosomiasis infections often have a benign course, and generally, should not prevent LT. The available literature was reviewed and a provisional screening protocol has been proposed.

Schistosoma mansoni infection in residents of a riverside community in Eastern Amazon.

Schistosoma mansoni worms cause a waterborne parasitic disease called schistosomiasis. It commonly affects individuals in lack of sanitation structure. In Brazil, Pará state has Belém as one of the worst sanitation-ranking places in 2023, where schistosomiasis transmission was already documented. This study reports the occurrence of schistosomiasis in residents of Ilha das Onças, an island next to Belém. Stool samples were obtained from participants over 2 years old, all residents from Furo do Rio Grande, one of the rivers on the island. The Kato-Katz technique was performed for parasite investigation in the stool samples. Each participant responded to a sociodemographic and clinical questionnaire. The residences were georeferenced for map designing. Three out of 263 participants were S. mansoni positive, all men, ages ranging from 19 to 41 years old, with low parasitic load. Malacological surveys were carried out, but no Biomphalaria snails were found. Risk factors for schistosomiasis establishment are present on the island, and the lack of sanitation makes it a potential risk area. Malacological surveys are highly encouraged as preventive measures, as well as health surveillance for riverside populations, generating data that will help health authorities in the management and planning of preventive control actions.

Geospatial analysis and prevalence of Schistosoma mansoni and soil-transmitted helminth infections in an endemic area in Eastern Brazilian Amazon.

OBJECTIVES: This study evaluated the occurrence of Schistosoma mansoni and soil-transmitted helminths in an endemic area in the Eastern Brazilian Amazon, analysing prevalence and spatial distribution. METHODS: The study was conducted in four localities of Primavera Municipality, in Pará state. Data was obtained from the Decit 40/2012 project and the participants were divided into five age range categories for evaluation: children, adolescents, young adults, adults and elderly individuals. For the diagnostic tests, Kato-Katz slides were prepared to detect S. mansoni and soil-transmitted helminths eggs. The spatial distribution map and the Kernel Density Estimation were performed to assess the presence and location of infections. RESULTS: Stool samples revealed the presence of hookworms, S. mansoni, Ascaris lumbricoides and Trichuris trichiura eggs. Mono-, bi- and poly-parasitic infections were observed, with a significant prevalence of hookworm monoparasitism. CONCLUSIONS: The high frequency of children infected with soil-transmitted helminths confirms their significance as an ongoing public health problem in the poorest municipalities of Brazil. The Geographic Information System plays a crucial role in environmental surveillance and in the control of epidemics and endemic diseases, enabling accurate assessment and informed decision-making for their control.

Cryptic population structure and transmission dynamics uncovered for Schistosoma mansoni populations by genetic analyses.

Patterns of diversity in pathogen genomes provide a window into the spatiotemporal spread of disease. In this study, we tested the hypothesis that Schistosoma mansoni parasites form genetic clusters that coincide with the communities of their human hosts. We also looked for genetic clustering of parasites at the sub-community level. Our data consists of 14 microsatellite DNA markers, typed from pooled DNA samples from [Formula: see text] infected individuals living in three Brazilian communities. We found a one-to-one correspondence between genetic clusters found by K-means cluster analysis and communities when [Formula: see text]. These clusters are also easily identified in a neighbor-joining tree and principal coordinates plots. K-means analysis with [Formula: see text] also reveals genetic clusters of parasites at the sub-community level. These sub-clusters also appear on the neighbor-joining tree and principal coordinates plots. A surprising finding is a genetic relationship between subgroups in widely separated human communities. This connection suggests the existence of common transmission sites that have wide influence. In summary, the genetic structure of S. mansoni in Brazil juxtaposes local isolation that is occasionally broken by long-range migration. Permanent eradication of schistosomes will require both local efforts and the identification of regional infection reservoirs.

Neurological impairment caused by Schistosoma mansoni systemic infection exhibits early features of idiopathic neurodegenerative disease.

Schistosomiasis, a neglected tropical disease caused by trematodes of the Schistosoma genus, affects over 250 million people around the world. This disease has been associated with learning and memory deficits in children, whereas reduced attention levels, impaired work capacity, and cognitive deficits have been observed in adults. Strongly correlated with poverty and lack of basic sanitary conditions, this chronic endemic infection is common in Africa, South America, and parts of Asia and contributes to inhibition of social development and low quality of life in affected areas. Nonetheless, studies on the mechanisms involved in the neurological impairment caused by schistosomiasis are scarce. Here, we used a murine model of infection with Schistosoma mansoni in which parasites do not invade the central nervous system to evaluate the consequences of systemic infection on neurologic function. We observed that systemic infection with S. mansoni led to astrocyte and microglia activation, expression of oxidative stress-induced transcription factor Nrf2, oxidative damage, Tau phosphorylation, and amyloid-ß peptide accumulation in the prefrontal cortex of infected animals. We also found impairment in spatial learning and memory as evaluated by the Morris water maze task. Administration of anthelmintic (praziquantel) and antioxidant (N-acetylcysteine plus deferoxamine) treatments was effective in inhibiting most of these phenotypes, and the combination of both treatments had a synergistic effect to prevent such changes. These data demonstrate new perspectives toward the understanding of the pathology and possible therapeutic approaches to counteract long-term effects of systemic schistosomiasis on brain function.

Optimization of Expression and Purification of Schistosoma mansoni Antigens in Fusion with Rhizavidin.

Schistosomiasis causes significant morbidity and mortality. Vaccine efforts to date indicate the need to increase the immunogenicity of Schistosoma antigens. The multiple antigen-presenting system, whereby proteins are genetically fused to rhizavidin and affinity linked to biotinylated templates, enables the generation of robust immune responses. The objective of this work was to express and purify the S. mansoni antigens, SmTSP-2 and SmCD59.2, in fusion with rhizavidin. The fusion with rhizavidin greatly decreased the expression level of rSmTSP-2, but not rSmCD59.2, and both were expressed in the insoluble fraction, requiring optimization of culture conditions. Evaluation of different E. coli strains and media showed that BL21-DE3 cultured in Terrific Broth provided the highest expression levels of both proteins. Investigation of a range of time and temperature of induction showed that E. coli strains expressing rRzv:SmTSP-2 and rRzv:SmCD59.2 showed the highest protein production at 23 °C for 15 h. Recombinant proteins were purified by a single step of affinity chromatography allowing isolation of these proteins in high concentration and purity. The optimization process increased final soluble protein yield of rRzv:SmTSP-2 by fourfold and rRzv:SmCD59.2 by tenfold, providing ~ 20 mg/L of each protein. Optimized fusion protein production will allow antigen use in biotin-rhizavidin affinity platforms.

FMRF-NH2 -related neuropeptides in Biomphalaria spp., intermediate hosts for schistosomiasis: Precursor organization and immunohistochemical localization.

Freshwater snails of the genus Biomphalaria serve as intermediate hosts for the digenetic trematode Schistosoma mansoni, the etiological agent for the most widespread form of intestinal schistosomiasis. As neuropeptide signaling in host snails can be altered by trematode infection, a neural transcriptomics approach was undertaken to identify peptide precursors in Biomphalaria glabrata, the major intermediate host for S. mansoni in the Western Hemisphere. Three transcripts that encode peptides belonging to the FMRF-NH2 -related peptide (FaRP) family were identified in B. glabrata. One transcript encoded a precursor polypeptide (Bgl-FaRP1; 292 amino acids) that included eight copies of the tetrapeptide FMRF-NH2 and single copies of FIRF-NH2 , FLRF-NH2 , and pQFYRI-NH2 . The second transcript encoded a precursor (Bgl-FaRP2; 347 amino acids) that comprised 14 copies of the heptapeptide GDPFLRF-NH2 and 1 copy of SKPYMRF-NH2 . The precursor encoded by the third transcript (Bgl-FaRP3; 287 amino acids) recapitulated Bgl-FaRP2 but lacked the full SKPYMRF-NH2 peptide. The three precursors shared a common signal peptide, suggesting a genomic organization described previously in gastropods. Immunohistochemical studies were performed on the nervous systems of B. glabrata and B. alexandrina, a major intermediate host for S. mansoni in Egypt. FMRF-NH2 -like immunoreactive (FMRF-NH2 -li) neurons were located in regions of the central nervous system associated with reproduction, feeding, and cardiorespiration. Antisera raised against non-FMRF-NH2 peptides present in the tetrapeptide and heptapeptide precursors labeled independent subsets of the FMRF-NH2 -li neurons. This study supports the participation of FMRF-NH2 -related neuropeptides in the regulation of vital physiological and behavioral systems that are altered by parasitism in Biomphalaria.

Effects of droughts on the larval digenean assemblage that parasitizes the snail Biomphalaria peregrina (Planorbidae): a 4-year study in a temporary pond from Argentina.

Temporary ponds are ecologically valuable habitats and useful sites for studies of diversity, ecology, evolution, and climate change. Organisms inhabiting these environments have developed strategies to ensure their survival. However, little is known about the temporal dynamics and strategies of parasites in these habitats. A 4-year study was carried out in a temporary pond to analyze the larval digenean assemblage in Biomphalaria peregrina, a potential vector of Schistosoma mansoni, a zoonosis of global importance. This environment had intermittent and irregular hydroperiods, and the overall prevalence showed differences throughout the years. After drought seasons, eight out of a total of nine species of digeneans were observed, and almost half of the snails were parasitized. In addition, six of the nine species of digeneans showed some type of abbreviation of their life cycles. These results suggest that digeneans have certain degree of plasticity in their life cycles in response to environmental changes in this pond, and the abbreviation of the digenean life cycle could be regarded as a parasite strategy to resist prolonged periods of desiccation.

Diagnostic comparison of stool exam and point-of-care circulating cathodic antigen (POC-CCA) test for schistosomiasis mansoni diagnosis in a high endemicity area in northeastern Brazil.

This study aimed to evaluate the performance of the point-of-care circulating cathodic antigen (POC-CCA) test in a highly endemic area in Brazil, comparing it to the Kato-Katz (KK) technique for sensitivity, specificity and the intensity of the reaction of the test in relation to the parasitic load. The community in Sergipe, Brazil, participated in the study, providing three stool samples, one of urine (POC-CCA) and fingers tick blood sample was tested by enzyme-linked immunosorbent assay (ELISA). Sensitivity, specificity, positive predictive value, negative predictive value, accuracy, kappa coefficient and Spearman's correlation were calculated for the POC-CCA test using the KK as the reference. The prevalence of schistosomiasis by KK testing was 48.82%; POC-CCA (t+) 66.14%; POC-CCA (t-) 45.24%. ELISA results showed 100% agreement in individuals with high and moderate eggs per gram (EPG). POC-CCA presented good diagnostic performance in individuals with medium and high EPG, but there were a high number of false negatives in individuals with low intensity infections. As observed, POC-CCA-filter test improves accuracy and sensitivity compared to a conventional test.

Detection of Schistosoma mansoni long non-coding RNAs in the infected C57BL/6 mouse liver.

Long non-coding RNAs (lncRNAs) perform several types of regulatory functions and have been recently explored in the genus Schistosoma. Although sequencing and bioinformatics approaches have demonstrated the presence of hundreds of lncRNAs and microRNAs (miRNAs) in this genus, information regarding their abundance, characteristics, and potential functions linked to Schistosoma mansoni biology and parasite-host interaction is limited. Our objectives in the present study were to verify whether 15 previously identified S. mansoni lncRNAs are detectable in the host liver. In addition, we assess whether these lncRNAs are present in the S. mansoni infective form and the stages inside the definitive host. The detection of these 15 S. mansoni lncRNAs and a long terminal repeat (LTR) retrotransposon Saci 4 was performed in the eggs, cercariae, and 3.5-h schistosomula. All lncRNAs were found to be expressed in these stages; some of the lncRNAs were found in the livers of the infected C57BL/6 mice. In conclusion, S. mansoni lncRNAs were detected in host livers and quantified. Furthermore, many of the lncRNAs analyzed showed differential expression in the larval stages, indicating that they play a stage-specific regulatory role.

Aspectos relacionados com a positividade para a esquistossomose: estudo transversal em área de baixa prevalência em Alagoas, 2020 / Aspectos relacionados con la positividad para la esquistosomiasis: un estudio transversal en un área de baja prevalencia en Alagoas, Brasil, 2020 / Aspects related to positivity for schistosomiasis: a cross-sectional study in a low prevalence area in Alagoas, Brazil, 2020

Objetivo: Analisar aspectos relacionados com a positividade para esquistossomose em área de baixa prevalência, no Brasil. Métodos: Estudo transversal, realizado no primeiro semestre de 2020, quando foram analisadas a proporção de positividade, em função do número de lâminas de Kato-Katz, o desempenho diagnóstico do teste e a estimação da positividade a partir dos dados do Sistema de Informação do Programa de Vigilância e Controle da Esquistossomose (SISPCE). Resultados: Foram analisadas 2.088 lâminas de 348 indivíduos, sendo a proporção de positividade de 11,8%, 26,7% e 31,0% para 1, 4 e 6 lâminas analisadas, respectivamente. Houve concordância excelente (índice Kappa = 0,91) na comparação entre as leituras de 4 e 6 lâminas. Foi estimada subnotificação de 2,1 vezes nos dados do SISPCE. Conclusão: Ampliar o número de lâminas aumentou a positividade do Kato-Katz, o que pode contribuir para maximizar o controle da doença enquanto problema de Saúde Pública.

Objetivo: Analizar aspectos relacionados con la positividad para esquistosomiasis en área de baja prevalencia en Brasil. Métodos: Estudio transversal, realizado en el primer semestre de 2020, donde se analizó la proporción de positividad según el número de portaobjetos de Kato-Katz, el rendimiento diagnóstico de la prueba y la creación de un factor de estimación de positividad a partir de los datos del Sistema de Información del Programa de Vigilancia y Control de la Esquistosomiasis (SISPCE). Resultados: Se analizaron 2.088 láminas de 348 individuos, con proporción de positividad del 11,8%, 26,7% y 31,0% para 1, 4 y 6 láminas analizadas, respectivamente. Hubo una excelente concordancia (Kappa = 0,91) en la comparación entre la lectura de 4 y 6 láminas. Se estimó un subregistro de 2,1 veces en los datos del SISPCE. Conclusión: Aumentar el número de muestras aumentó la positividad de Kato-Katz, lo que puede contribuir a maximizar el control de la enfermedad como problema de Salud Pública.

Objective: To analyze aspects related to schistosomiasis positivity in an area of low prevalence in Brazil. Methods: This was a cross-sectional study, carried out in the first half of 2020, where we analyzed the proportion of positivity, according to the number of Kato-Katz slides, the diagnostic performance of the test and positivity estimates based on data from the Schistosomiasis Surveillance and Control Program Information System (SISPCE). Results: 2,088 slides from 348 individuals were analyzed, with proportion of positivity of 11.8%, 26.7% and 31.0% for 1, 4 and 6 slides analyzed, respectively. There was excellent agreement (Kappa = 0.91) between the readings of 4 and 6 slides. The SISPCE data was estimated to be underreported by up to 2.1 times. Conclusion: Increasing the number of slides increased Kato-Katz positivity, which can contribute to maximizing the control of the disease as a Public Health problem.

Identification of Schistosoma mansoni Infection in a Nonhuman Primate from St. Kitts More than 50 Years after Interruption of Human Transmission.

Transmission of Schistosoma mansoni was interrupted on St. Kitts, a Caribbean island, in the 1950s. With no reported cases since that time and most Biomphalaria spp. snail populations eliminated based on surveys in the 1970s, S. mansoni has been considered eliminated on St. Kitts. In 2019, S. mansoni eggs were found in an African green monkey (Chlorocebus aethiops sabaeus) that originated from St. Kitts. Nonhuman primate (NHP) infections have been considered incidental to human infections, with infections in NHPs resolving with the elimination of S. mansoni in the human population. An NHP with S. mansoni infection suggests that the NHP may be able to maintain a reservoir sylvatic cycle. Alternatively, S. mansoni transmission was not eliminated or S. mansoni has been reintroduced to St. Kitts. The occurrence of an infected NHP from St. Kitts supports the need for continuous monitoring in areas where S. mansoni is considered eliminated.

The use of the circulating cathodic antigen (CCA) urine cassette assay for the diagnosis and assessment of cure of Schistosoma mansoni infections in an endemic area of the Amazon region.

INTRODUCTION: Schistosomiasis is a poverty-related disease that affects people in 78 countries worldwide. This study aimed to evaluate the point-of-care circulating cathodic antigen (POC-CCA) test performance using sensitive parasitological methods as a reference standard (RS) in individuals before and after treatment. METHODS: The RS was established by combining the results of 16 Kato-Katz slides and the Helmintex® method. Positivity rates of the POC-CCA test and Kato-Katz and Helmintex® methods were calculated before treatment and 30 days afterward. Furthermore, the sensitivity, specificity, accuracy, and kappa coefficient before treatment were determined by comparing the methods. The cure rate was defined 30 days after treatment. RESULTS: Among the 217 participants, the RS detected a total of 63 (29.0%) positive individuals. The POC-CCA test identified 79 (36.4%) infections. The evaluation of POC-CCA test performance in relation to the RS revealed a sensitivity of 61.9%, specificity of 74.0%, accuracy of 70.5%, and kappa coefficient of 0.33. Out of the 53 remaining participants after treatment, a total of 45 (81.1%) showed egg negative results, and 8 (18.9%) were egg positive according to the RS. A total of 5 (9.4%) egg-positive and 37 (69.8%) egg-negative individuals were positive by the POC-CCA test. CONCLUSIONS: Our data show that the POC-CCA test has potential as an auxiliary tool for the diagnosis of Schistosoma mansoni infection, yielding better results than 16 Kato-Katz slides from three different stool samples. However, the immunochromatographic test lacks sufficient specificity and sensitivity for verifying the cure rate after treatment.

A new diagnostic strategy which uses a luminol-H2O2 system to detect helminth eggs in fecal sediments processed by the Helmintex method.

Schistosomiasis remains a serious public health problem in tropical regions, affecting more than 250 million people. Sensitive diagnostic methods represent key tools for disease elimination, in particular in areas with low endemicity. Advances in the use of luminol-based chemiluminescent techniques have enabled greater sensitivity and speed in obtaining results in different diagnostic settings. In this study, we developed a luminol-H2O2 chemiluminescence (CL) method to detect Schistosoma mansoni eggs in human fecal sediments processed by the Helmintex (HTX) method. After S. mansoni eggs were incubated with a solution of luminol-H2O2 the light emission was detected and measured by spectrophotometry at 431 nm for 5 min, using detection and counts of eggs by bright field optical microscopy as a reference. CL intensity was found to correlate with different sources and numbers of eggs. Furthermore, our results showed that the CL method can distinguish positive from negative samples with 100% sensitivity and 71% specificity. To our knowledge, this is the first study to report the use of CL for the diagnosis of helminths from fecal samples. The combination of the HTX method with CL represents an important advance in providing a reference method with the highest standards of sensitivity.

Analysis and spatial distribution of schistosomiasis mansoni in a historically endemic area of northeastern Brazil.

OBJECTIVE: To perform a descriptive analysis of the activities of the Schistosomiasis Control Program, as well as the spatial distribution of the condition in the state of Alagoas, Brazil, for the period from 2007 to 2016. METHODS: Descriptive ecological study. Data from positive human cases and operational data were collected in the Information System of the Schistosomiasis Control Program, and data for spatial analysis were collected on the website of the Brazilian Institute of Geography and Statistics. An analysis of spatial autocorrelation (Moran statistics) was performed, where a spatial pattern was established, which showed the Q1 and Q2 patterns to be the most important, and Q3 and Q4 representing transition areas. RESULTS: In the years under study, at least 85% (n = 60/70) of the municipalities carried out the activities recommended by the PCE (Schistosomiasis Control Program). Alagoas presented an average positivity rate of 7.1%, which is very high compared to the prevalence of 3.3% at the last national schistosomiasis survey conducted between 2010 and 2015. Moran's statistics showed 22/70 municipalities forming a Q1 cluster, of high/high pattern, and 32/70 municipalities forming a Q2 cluster, of low/low pattern, with the others in a transition area. Moran Map data, however, showed only 7/70 municipalities in the endemic area with a spatial autocorrelation, with these municipalities having the Mundau River as a common element. CONCLUSION: Schistosomiasis mansoni is of great importance for public health in Alagoas and that the use of spatial analysis can identify priority areas for preventive and control measures against schistosomiasis mansoni.

OBJECTIF: Effectuer une analyse descriptive des activités du programme de lutte contre la schistosomiase, ainsi que la distribution spatiale de la condition dans l'Etat d'Alagoas, au Brésil, pour la période de 2007 à 2016. MÉTHODES: Etude écologique descriptive. Les données sur les cas humains positifs et les données opérationnelles ont été collectées dans le système d'information du programme de lutte contre la schistosomiase et les données pour l'analyse spatiale ont été collectées du site Web de l'Institut brésilien de géographie et des statistiques. Une analyse de l'autocorrélation spatiale (statistiques de Moran) a été réalisée, où un modèle spatial a été établi, qui a montré que les modèles Q1 et Q2 étaient les plus importants, Q3 et Q4 représentant les zones de transition. RÉSULTATS: Au cours de toutes les années étudiées, au moins 85% (n = 60/70) des municipalités ont réalisé les activités recommandées par le PCE (Program de Contrôle de la Schistosomiase). En ce qui concerne le taux de positivité, Alagoas présentait un taux de positivité moyen de 7,1% pour la période étudiée, considéré comme très élevé par rapport aux données de la dernière enquête nationale sur la schistosomiase réalisée entre 2010 et 2015, qui montrait l'état avec une prévalence de 3,3%. Les statistiques de Moran ont montré 22/70 municipalités formant un regroupement Q1, de modèle élevé/élevé, et 32/70 municipalités formant un regroupement Q2, de modèle faible/faible, avec les autres dans une zone de transition. Les données de la carte Moran, cependant, ne montraient que 7/70 municipalités de la zone endémique avec une autocorrélation spatiale, ces municipalités ayant la rivière Mundau comme élément commun. CONCLUSION: Les données présentées ici montrent que la schistosomiase mansonnienne est d'une grande ampleur pour la santé publique à Alagoas et que l'utilisation de l'analyse spatiale peut identifier les domaines prioritaires pour l'adoption de mesures de prévention et de contrôle de la schistosomiase mansonnienne.

Accuracy of real-time polymerase chain reaction to detect Schistosoma mansoni - infected individuals from an endemic area with low parasite loads.

Due to the efforts to control schistosomiasis transmission in tropical countries, a large proportion of individuals from endemic areas present low parasite loads, which hinders diagnosis of intestinal schistosomiasis by the Kato-Katz (KK) method. Therefore, the development of more sensitive diagnostic methods is essential for efficient control measures. The aim was to evaluate the accuracy of a real-time polymerase chain reaction (RT-PCR) to detect Schistosoma mansoni DNA in fecal samples of individuals with low parasite loads. A cross-sectional population-based study was conducted in a rural community (n = 257) in Brazil. POC-CCA® was performed in urine and feces were used for RT-PCR. In addition, fecal exams were completed by 18 KK slides, saline gradient and Helmintex techniques. The combined results of the three parasitological tests detected schistosome eggs in 118 participants (45.9%) and composed the consolidated reference standard (CRS). By RT-PCR, 117 out of 215 tested samples were positive, showing 91.4% sensitivity, 80.2% specificity and good concordance with the CRS (kappa = 0.71). RT-PCR identified 86.9% of the individuals eliminating less than 12 eggs/g of feces, demonstrating much better performance than POC-CCA® (50.8%). Our results showed that RT-PCR is a valuable alternative for the diagnosis of intestinal schistosomiasis in individuals with very low parasite loads.

Circulating Anodic Antigen (CAA): A Highly Sensitive Diagnostic Biomarker to Detect Active Schistosoma Infections-Improvement and Use during SCORE.

The Schistosomiasis Consortium for Operational Research and Evaluation (SCORE) was funded in 2008 to conduct research that would support country schistosomiasis control programs. As schistosomiasis prevalence decreases in many places and elimination is increasingly within reach, a sensitive and specific test to detect infection with Schistosoma mansoni and Schistosoma haematobium has become a pressing need. After obtaining broad input, SCORE supported Leiden University Medical Center (LUMC) to modify the serum-based antigen assay for use with urine, simplify the assay, and improve its sensitivity. The urine assay eventually contributed to several of the larger SCORE studies. For example, in Zanzibar, we demonstrated that urine filtration, the standard parasite egg detection diagnostic test for S. haematobium, greatly underestimated prevalence in low-prevalence settings. In Burundi and Rwanda, the circulating anodic antigen (CAA) assay provided critical information about the limitations of the stool-based Kato-Katz parasite egg-detection assay for S. mansoni in low-prevalence settings. Other SCORE-supported CAA work demonstrated that frozen, banked urine specimens yielded similar results to fresh ones; pooling of specimens may be a useful, cost-effective approach for surveillance in some settings; and the assay can be performed in local laboratories equipped with adequate centrifuge capacity. These improvements in the assay continue to be of use to researchers around the world. However, additional work will be needed if widespread dissemination of the CAA assay is to occur, for example, by building capacity in places besides LUMC and commercialization of the assay. Here, we review the evolution of the CAA assay format during the SCORE period with emphasis on urine-based applications.

SCHISTOSOMAL MYELORADICULOPATHY IN A NON-ENDEMIC AREA.

OBJECTIVE: To report a schistosomal myeloradiculopathy case in a non-endemic area. CASE DESCRIPTION: A previously healthy 11-year-old boy, stricken by an acute loss of strength on his lower limbs, followed by a loss of strength on his upper limbs and upper body, associated with altered sensitivity of the vesical globe formation. The patient's cerebrospinal fluid analysis showed eosinophilic meningitis, in addition to peripheral eosinophilia. The investigation resulted in a positive serology for Schistosoma mansoni. The treatment included steroids and praziquantel 60mg/kg, with a new dose after a month, as well as physical therapy for rehabilitation. The patient evolved with clinical improvement in the neurological exam, with a medullary section initially at C6, but now at T6. The patient is kept at prednisolone use (30mg/day) and longterm urinary catheter dependence. COMMENTS: The schistosomiasis is endemic in many regions of Brazil; however, it has low incidence in the south of the country. Among its main manifestations, the schistosomal myeloradiculopathy is the most severe ectopic form of the disease, and should be suspected in patients with low back pain, strength and/or sensibility disorder of the lower limbs or urinary tract's disturbance. Early diagnosis and treatment should be done in order to reduce severe neurological sequelae. Treatment includes schistosomiasis drugs, corticosteroids and/or surgery.

A Real-Time PCR Assay for the Diagnosis of Intestinal Schistosomiasis and Cure Assessment After the Treatment of Individuals With Low Parasite Burden.

The laboratorial diagnosis of the intestinal schistosomiasis is always performed using Kato-Katz technique. However, this technique presents low sensitivity for diagnosis of individuals with low parasite burden, which constitutes the majority in low endemicity Brazilian locations for the disease. The objective of this study was developed and to validate a real-time PCR assay (qPCR) targeting 121 bp sequence to detect Schistosoma spp. DNA for the diagnosis of intestinal schistosomiasis and a sequence of the human ß-actin gene as internal control. Firstly, the qPCR was standardized and next it was evaluated for diagnosis and cure assessment of intestinal schistosomiasis in the resident individuals in Tabuas and Estreito de Miralta, two locations in Brazil endemic for intestinal schistosomiasis. The qPCR assay results were compared with those of the Kato-Katz (KK) test, examining 2 or 24 slides, Saline Gradient (SG) and "reference test" (24 KK slides + SG). The cure assessment was measured by these diagnostic techniques at 30, 90, and 180 days post-treatment. In Tabuas, the positivity rates obtained by the qPCR was 30.4% (45/148) and by "reference test" was of 31.0% (46/148), with no statistical difference (p = 0.91). The presumed cure rates at 30, 90, and 180 days post-treatment were 100, 94.4, and 78.4% by the analysis of 24 KK slides, 100, 94.4, and 78.4% by the SG, and 100, 83.3, and 62.1% by the qPCR assay. In Estreito de Miralta, the positivity obtained by qPCR was 18.3% (26/142) and with "reference test" was 24.6% (35/142), with no statistical difference (p = 0.20). The presumed cure rates were 93.3, 96.9, and 96.5% by the KK, 93.3, 96.9, and 100% by the SG, and 93.3, 93.9, and 96.5% by the qPCR at 30, 90, and 180 days post-treatment, respectively. This study showed that the diagnostic techniques presented different performance in the populations from the two districts (Tabuas and Estreito de Miralta) and reinforces the need of combining techniques to improve diagnosis accuracy, increasing the detection of individuals with low parasite burden. This combination of techniques consists an important strategy for controlling the disease transmission.