Culture-free genome-wide locus sequence typing (GLST) provides new perspectives on Trypanosoma cruzi dispersal and infection complexity.

Analysis of genetic polymorphism is a powerful tool for epidemiological surveillance and research. Powerful inference from pathogen genetic variation, however, is often restrained by limited access to representative target DNA, especially in the study of obligate parasitic species for which ex vivo culture is resource-intensive or bias-prone. Modern sequence capture methods enable pathogen genetic variation to be analyzed directly from host/vector material but are often too complex and expensive for resource-poor settings where infectious diseases prevail. This study proposes a simple, cost-effective 'genome-wide locus sequence typing' (GLST) tool based on massive parallel amplification of information hotspots throughout the target pathogen genome. The multiplexed polymerase chain reaction amplifies hundreds of different, user-defined genetic targets in a single reaction tube, and subsequent agarose gel-based clean-up and barcoding completes library preparation at under 4 USD per sample. Our study generates a flexible GLST primer panel design workflow for Trypanosoma cruzi, the parasitic agent of Chagas disease. We successfully apply our 203-target GLST panel to direct, culture-free metagenomic extracts from triatomine vectors containing a minimum of 3.69 pg/µl T. cruzi DNA and further elaborate on method performance by sequencing GLST libraries from T. cruzi reference clones representing discrete typing units (DTUs) TcI, TcIII, TcIV, TcV and TcVI. The 780 SNP sites we identify in the sample set repeatably distinguish parasites infecting sympatric vectors and detect correlations between genetic and geographic distances at regional (< 150 km) as well as continental scales. The markers also clearly separate TcI, TcIII, TcIV and TcV + TcVI and appear to distinguish multiclonal infections within TcI. We discuss the advantages, limitations and prospects of our method across a spectrum of epidemiological research.

Strong effect of Ecuador's conditional cash transfer program on childhood mortality from poverty-related diseases: a nationwide analysis.

BACKGROUND: The mortality rate in children under 5 years old (U5MR) has decreased considerably in Ecuador in the last decade; however, thousands of children continue to die from causes related to poverty. A social program known as Bono de Desarrollo Humano (BDH) was created to guarantee a minimum level of consumption for families and to reduce chronic malnutrition and preventable childhood diseases. We sought to evaluate the effect of the BDH program on mortality of children younger than 5 years, particularly from malnutrition, diarrheal diseases, and lower respiratory tract infections. METHODS: Mortality rates and BDH coverage from 2009 to 2014 were evaluated from the 144 (of 222) Ecuadorian counties with intermediate and high quality of vital information. A multivariable regression analyses for panel data was conducted by using a negative binomial regression model with fixed effects, adjusted for all relevant demographic and socioeconomic covariates. RESULTS: Our research shows that for each 1% increase in BDH county coverage there would be a decrease in U5MR from malnutrition of 3% (RR 0.971, 95% CI 0.953-0.989). An effect of BDH county coverage on mortality resulting from respiratory infections was also observed (RR 0.992, 95% CI 0.984-0.999). The BDH also reduced hospitalization rates in children younger than 5 years, overall and for diarrhea. CONCLUSIONS: A conditional cash transfer program such as BDH could contribute to the reduction of mortality due to causes related to poverty, such as malnutrition and respiratory infections. The coverage should be maintained -or increased in a period of economic crisis- and its implementation strengthened.

Home improvement and system-based health promotion for sustainable prevention of Chagas disease: A qualitative study.

BACKGROUND: Human transmission of Chagas disease (CD) most commonly occurs in domiciliary spaces where triatomines remain hidden to feed on blood sources during inhabitants' sleep. Similar to other neglected tropical diseases (NTDs), sustainable control of CD requires attention to the structural conditions of life of populations at risk, in this case, the conditions of their living environments. Considering socio-cultural and political dynamics involved in dwellings' construction, this study aimed to explore social factors that contribute or limit sustainability of CD's prevention models focused on home improvement. METHODS AND MAIN FINDINGS: Using Healthy Homes for Healthy Living (HHHL)-a health promotion strategy focused on improvement of living environments and system-based health promotion-as a reference, a qualitative study was conducted. Research participants were selected from three rural communities of a CD endemic region in southern Ecuador involved in HHHL's refurbishment and reconstruction interventions between 2013 and 2016. Folowing an ethnographic approach, data were collected through interviews, participant observation, informal conversations and document analysis. Our results indicate that the HHHL model addressed risk factors for CD at the household level, while simultaneously promoting wellbeing at emotional, economic and social levels in local communities. We argue that sustainability of the CD prevention model proposed by HHHL is enhanced by the confluence of three factors: systemic improvement of families' quality of life, perceived usefulness of control measures, and flexibility to adapt to emerging dynamics of the context. CONCLUSION: HHHL's proposed home improvement, facilitated through system-based rather than disease specific health promotion processes, enhances agency in populations at risk and facilitates community partnerships forged around CD prevention. Although an independent analysis of cost-effectiveness is recommended, structural poverty experienced by local families is still the most important factor to consider when evaluating the sustainability and scalability of this model.

A model of insect control with imperfect treatment

Insecticide spraying of housing units is an important control measure for vector-borne infections such as Chagas disease. However, some vectors may survive treatment, due to imperfect spraying by the operator or because they hide deep in the cracks or other places, and re-emerge in the same unit when the effect of the insecticide wears off. While several mathematical models of this phenomenon have been previously described and studied in the literature, the model presented here is more basic than existing ones. Thus it is more amenable to mathematical analysis, which is carried out here. In particular, we demonstrate that an initially very high spraying rate may push the system into a region of the state space with low endemic levels of infestation that can be maintained in the long run at relatively moderate cost, while in the absence of an aggressive initial intervention the same average cost would only allow a much less significant reduction in long-term infestation levels.

Models of Disease Vector Control: When Can Aggressive Initial Intervention Lower Long-Term Cost?

Insecticide spraying of housing units is an important control measure for vector-borne infections such as Chagas disease. As vectors may invade both from other infested houses and sylvatic areas and as the effectiveness of insecticide wears off over time, the dynamics of (re)infestations can be approximated by [Formula: see text]-type models with a reservoir, where housing units are treated as hosts, and insecticide spraying corresponds to removal of hosts. Here, we investigate three ODE-based models of this type. We describe a dual-rate effect where an initially very high spraying rate can push the system into a region of the state space with low endemic levels of infestation that can be maintained in the long run at relatively moderate cost, while in the absence of an aggressive initial intervention the same average cost would only allow a much less significant reduction in long-term infestation levels. We determine some sufficient and some necessary conditions under which this effect occurs and show that it is robust in models that incorporate some heterogeneity in the relevant properties of housing units.

2b-RAD genotyping for population genomic studies of Chagas disease vectors: Rhodnius ecuadoriensis in Ecuador.

BACKGROUND: Rhodnius ecuadoriensis is the main triatomine vector of Chagas disease, American trypanosomiasis, in Southern Ecuador and Northern Peru. Genomic approaches and next generation sequencing technologies have become powerful tools for investigating population diversity and structure which is a key consideration for vector control. Here we assess the effectiveness of three different 2b restriction site-associated DNA (2b-RAD) genotyping strategies in R. ecuadoriensis to provide sufficient genomic resolution to tease apart microevolutionary processes and undertake some pilot population genomic analyses. METHODOLOGY/PRINCIPAL FINDINGS: The 2b-RAD protocol was carried out in-house at a non-specialized laboratory using 20 R. ecuadoriensis adults collected from the central coast and southern Andean region of Ecuador, from June 2006 to July 2013. 2b-RAD sequencing data was performed on an Illumina MiSeq instrument and analyzed with the STACKS de novo pipeline for loci assembly and Single Nucleotide Polymorphism (SNP) discovery. Preliminary population genomic analyses (global AMOVA and Bayesian clustering) were implemented. Our results showed that the 2b-RAD genotyping protocol is effective for R. ecuadoriensis and likely for other triatomine species. However, only BcgI and CspCI restriction enzymes provided a number of markers suitable for population genomic analysis at the read depth we generated. Our preliminary genomic analyses detected a signal of genetic structuring across the study area. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that 2b-RAD genotyping is both a cost effective and methodologically simple approach for generating high resolution genomic data for Chagas disease vectors with the power to distinguish between different vector populations at epidemiologically relevant scales. As such, 2b-RAD represents a powerful tool in the hands of medical entomologists with limited access to specialized molecular biological equipment.

Factores sociales asociados con la utilización de los servicios de atención prenatal en Ecuador / Social factors associated with use of prenatal care in Ecuador

Objetivos. La atención prenatal es uno de los pilares de la salud pública y permite el acceso a intervenciones tales como la prevención de la transmisión materno-infantil del VIH y de la sífilis congénita. Este artículo tiene como objetivo describir los factores sociales asociados con la utilización de los servicios de atención prenatal en Ecuador. Métodos. Entre 2011 y 2012, se realizó un análisis de la información procedente de las historias clínicas y de la entrevista a las participantes, que integraron una muestra probabilística a nivel nacional de 5 998 mujeres atendidas por parto o aborto en 15 servicios sanitarios en Ecuador con el objetivo de estimar la prevalencia de VIH, sífilis, enfermedad de Chagas y la cobertura de atención prenatal. Resultados. El estudio mostró que 94,1% de las mujeres había acudido a algún control prenatal, pero la asistencia al menos a cuatro controles fue 73,1%. Se encontró que el menor nivel educativo, el mayor número de embarazos, la ocupación en el sector agrícola o ganadero y la pertenencia a los grupos étnicos indígena, afroecuatoriano u otros minoritarios fueron factores asociados con la falta de uso (ningún control prenatal) o al uso inadecuado de la atención prenatal (menos de cuatro controles o primer control después de las 20 semanas de gestación) en Ecuador. Conclusiones. Estos resultados apuntan a la persistencia de desigualdades marcadas en el acceso y en la utilización de servicios de atención prenatal atribuibles a factores socioeconómicos y a la necesidad de fortalecer las estrategias para su abordaje para alcanzar la meta de la cobertura universal de atención prenatal.

Objectives. Prenatal care is a pillar of public health, enabling access to interventions including prevention of mother-to-child transmission of HIV and congenital syphilis. This paper describes social factors related to use of prenatal care in Ecuador. Methods. In 2011 and 2012, participant clinical history and interview information was analyzed from a national probability sample of 5 998 women presenting for delivery or miscarriage services in 15 healthcare facilities in Ecuador, to estimate prevalence of HIV, syphilis, and Chagas disease, and prenatal care coverage. Results. The study found that 94.1% of women had attended at least one prenatal visit, but that attendance at no less than four visits was 73.1%. Furthermore, lower educational level, greater number of pregnancies, occupation in the agriculture or livestock sector, and membership in ethnic indigenous, Afro-Ecuadorian, or other minority groups were factors associated with lack of use (no prenatal visits) or insufficient use of prenatal care (fewer than four visits or first visit at >20 weeks gestation) in Ecuador. Conclusions. These results point to persistence of marked inequalities in access to and use of prenatal health services attributable to socioeconomic factors and to the need to strengthen strategies to address them, to reach the goal of universal prenatal care coverage.

Factores sociales asociados con la utilización de los servicios de atención prenatal en Ecuador / Social factors associated with use of prenatal care in Ecuador

RESUMEN Objetivos La atención prenatal es uno de los pilares de la salud pública y permite el acceso a intervenciones tales como la prevención de la transmisión materno-infantil del VIH y de la sífilis congénita. Este artículo tiene como objetivo describir los factores sociales asociados con la utilización de los servicios de atención prenatal en Ecuador. Métodos Entre 2011 y 2012, se realizó un análisis de la información procedente de las historias clínicas y de la entrevista a las participantes, que integraron una muestra probabilística a nivel nacional de 5 998 mujeres atendidas por parto o aborto en 15 servicios sanitarios en Ecuador con el objetivo de estimar la prevalencia de VIH, sífilis, enfermedad de Chagas y la cobertura de atención prenatal. Resultados El estudio mostró que 94,1% de las mujeres había acudido a algún control prenatal, pero la asistencia al menos a cuatro controles fue 73,1%. Se encontró que el menor nivel educativo, el mayor número de embarazos, la ocupación en el sector agrícola o ganadero y la pertenencia a los grupos étnicos indígena, afroecuatoriano u otros minoritarios fueron factores asociados con la falta de uso (ningún control prenatal) o al uso inadecuado de la atención prenatal (menos de cuatro controles o primer control después de las 20 semanas de gestación) en Ecuador. Conclusiones Estos resultados apuntan a la persistencia de desigualdades marcadas en el acceso y en la utilización de servicios de atención prenatal atribuibles a factores socioeconómicos y a la necesidad de fortalecer las estrategias para su abordaje para alcanzar la meta de la cobertura universal de atención prenatal.(AU)

ABSTRACT Objectives Prenatal care is a pillar of public health, enabling access to interventions including prevention of mother-to-child transmission of HIV and congenital syphilis. This paper describes social factors related to use of prenatal care in Ecuador. Methods In 2011 and 2012, participant clinical history and interview information was analyzed from a national probability sample of 5 998 women presenting for delivery or miscarriage services in 15 healthcare facilities in Ecuador, to estimate prevalence of HIV, syphilis, and Chagas disease, and prenatal care coverage. Results The study found that 94.1% of women had attended at least one prenatal visit, but that attendance at no less than four visits was 73.1%. Furthermore, lower educational level, greater number of pregnancies, occupation in the agriculture or livestock sector, and membership in ethnic indigenous, Afro-Ecuadorian, or other minority groups were factors associated with lack of use (no prenatal visits) or insufficient use of prenatal care (fewer than four visits or first visit at >20 weeks gestation) in Ecuador. Conclusions These results point to persistence of marked inequalities in access to and use of prenatal health services attributable to socioeconomic factors and to the need to strengthen strategies to address them, to reach the goal of universal prenatal care coverage.(AU)

[Social factors associated with use of prenatal care in Ecuador]. / Factores sociales asociados con la utilización de los servicios de atención prenatal en Ecuador.

OBJECTIVES: Prenatal care is a pillar of public health, enabling access to interventions including prevention of mother-to-child transmission of HIV and congenital syphilis. This paper describes social factors related to use of prenatal care in Ecuador. METHODS: In 2011 and 2012, participant clinical history and interview information was analyzed from a national probability sample of 5 998 women presenting for delivery or miscarriage services in 15 healthcare facilities in Ecuador, to estimate prevalence of HIV, syphilis, and Chagas disease, and prenatal care coverage. RESULTS: The study found that 94.1% of women had attended at least one prenatal visit, but that attendance at no less than four visits was 73.1%. Furthermore, lower educational level, greater number of pregnancies, occupation in the agriculture or livestock sector, and membership in ethnic indigenous, Afro-Ecuadorian, or other minority groups were factors associated with lack of use (no prenatal visits) or insufficient use of prenatal care (fewer than four visits or first visit at >20 weeks gestation) in Ecuador. CONCLUSIONS: These results point to persistence of marked inequalities in access to and use of prenatal health services attributable to socioeconomic factors and to the need to strengthen strategies to address them, to reach the goal of universal prenatal care coverage.