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Los casos de la enfermedad de Chagas aguda (ECA) se reportan en forma esporádica en el Perú. En este artículo se describe las características clínicas y epidemiológicas de ocho nuevos casos de ECA detectados por el sistema de vigilancia en la cuenca amazónica, entre 2009 y 2016. La media de la edad fue 12,7 ± 13,7 años, rango de 1 a 44 años y 4/8 casos fueron varones. Un caso se asoció a enfermedad diarreica aguda, otro a leptospirosis y dos a infección del tracto urinario. La tasa de letalidad global fue 12,5% (1/8). La detección tardía es una característica frecuente relacionada a una escasa sospecha diagnóstica en pacientes con historial de fiebre. Se identificó el linaje TcI y TcIV del agente etiológico de la enfermedad de Chagas. Se reporta ocho nuevos casos de ECA, de los cuales siete fueron menores de edad.

Acute Chagas disease (ACD) cases are reported sporadically in Peru. In this report we describe the clinical and epidemiological characteristics of eight new ACD cases detected by the surveillance system in the Amazon basin, between 2009 and 2016. The average age was 12,7 ±13.7 years, range between 1 to 44 years and 4/8 cases were men. One case was associated with acute diarrheal disease, another with Leptospirosis, and two with urinary tract infection. The global case fatality-rate was 12.5% ​​(1/8). Late detection is a frequent characteristic related with low diagnostic suspicion in patients with a history of fever. The TcI and TcIV lineage was identified as the etiological agent of Chagas disease. Eight new cases of ACDs are reported, of which seven were children.Los casos de la enfermedad de Chagas aguda (ECA) se reportan en forma esporádica en el Perú. En este artículo se describe las características clínicas y epidemiológicas de ocho nuevos casos de ECA detectados por el sistema de vigilancia en la cuenca amazónica, entre 2009 y 2016. La media de la edad fue 12,7 ± 13,7 años, rango de 1 a 44 años y 4/8 casos fueron varones. Un caso se asoció a enfermedad diarreica aguda, otro a leptospirosis y dos a infección del tracto urinario. La tasa de letalidad global fue 12,5% (1/8). La detección tardía es una característica frecuente relacionada a una escasa sospecha diagnóstica en pacientes con historial de fiebre. Se identificó el linaje TcI y TcIV del agente etiológico de la enfermedad de Chagas. Se reporta ocho nuevos casos de ECA, de los cuales siete fueron menores de edad.

Anthropogenic environmental alterations such as urbanization can threaten native populations as well as create novel environments that allow human pests and pathogens to thrive. As the number and size of urban environments increase globally, it is more important than ever to understand the dispersal dynamics of hosts, vectors and pathogens of zoonotic disease systems. For example, a protozoan parasite and the causative agent of Chagas disease in humans, Trypanosoma cruzi, recently colonized and spread through the city of Arequipa, Peru. We used population genomic and phylogenomic tools to analyze whole genomes of 123 T. cruzi isolates derived from vectors and non-human mammals throughout Arequipa to determine patterns of T. cruzi dispersal. The data show significant population genetic structure within city blocks—parasites in the same block tend to be very closely related—but no population structure among blocks within districts—parasites in neighboring blocks are no more closely related to one another than to parasites in distant districts. These data suggest that T. cruzi dispersal within a block occurs regularly and that occasional long-range dispersal events allow the establishment of new T. cruzi populations in distant blocks. Movement of domestic animals may be the primary mechanism of inter-block and inter-district T. cruzi dispersal., Author summary Urbanization creates environments that are ideal for some human pests and pathogens. As the number and size of urban environments increases globally, it is becoming vital to understand how human disease-causing pathogens, their vectors, and their non-human hosts disperse through urban landscapes. Here we study a population of Trypanosoma cruzi–the protozoan parasite and causative agent of Chagas disease in humans–that recently colonized the city of Arequipa, Peru. We use population genomic and phylogenomic tools to understand how this parasite population dispersed through the city to achieve its current distribution and abundance. We show that T. cruzi collected from the same city block tend to be very closely related, while those from neighboring blocks are often as distantly related as those from blocks in distant districts. The data suggest that vectors facilitate frequent within-block dispersal of the parasite, while domestic animal movement may facilitate the relatively infrequent inter-block and inter-district dispersal.

Guinea pigs are important reservoirs of Trypanosoma cruzi , the causative parasite of Chagas disease, and in the Southern Cone of South America, transmission is mediated mainly by the vector Triatoma infestans . Interestingly, colonies of Triatoma infestans captured from guinea pig corrals sporadically have infection prevalence rates above 80%. Such high values are not consistent with the relatively short 7–8 week parasitemic period that has been reported for guinea pigs in the literature. We experimentally measured the infectious periods of a group of T. cruzi -infected guinea pigs by performing xenodiagnosis and direct microscopy each week for one year. Another group of infected guinea pigs received only direct microscopy to control for the effect that inoculation by triatomine saliva may have on parasitemia in the host. We observed infectious periods longer than those previously reported in a number of guinea pigs from both the xenodiagnosis and control groups. While some guinea pigs were infectious for a short time, other “super-shedders” were parasitemic up to 22 weeks after infection, and/or positive by xenodiagnosis for a year after infection. This heterogeneity in infectiousness has strong implications for T. cruzi transmission dynamics and control, as super-shedder guinea pigs may play a disproportionate role in pathogen spread.

Changing environmental conditions, including those caused by human activities, reshape biological communities through both loss of native species and establishment of non-native species in the altered habitats. Dynamic interactions with the abiotic environment impact both immigration and initial establishment of non-native species into these altered habitats. The repeated emergence of disease systems in urban areas worldwide highlights the importance of understanding how dynamic migratory processes affect the current and future distribution and abundance of pathogens in urban environments. In this study, we examine the pattern of invasion of Trypanosoma cruzi-the causative agent of human Chagas disease-in the city of Arequipa, Peru. Phylogenetic analyses of 136 T. cruzi isolates from Arequipa and other South American locations suggest that only one T. cruzi lineage established a population in Arequipa as all T. cruzi isolated from vectors in Arequipa form a recent monophyletic group within the broader South American phylogeny. We discuss several hypotheses that may explain the limited number of established T. cruzi lineages despite multiple introductions of the parasite.

Background Sexual reproduction provides an evolutionary advantageous mechanism that combines favorable mutations that have arisen in separate lineages into the same individual. This advantage is especially pronounced in microparasites as allelic reassortment among individuals caused by sexual reproduction promotes allelic diversity at immune evasion genes within individuals which is often essential to evade host immune systems. Despite these advantages, many eukaryotic microparasites exhibit highly-clonal population structures suggesting that genetic exchange through sexual reproduction is rare. Evidence supporting clonality is particularly convincing in the causative agent of Chagas disease, Trypanosoma cruzi, despite equally convincing evidence of the capacity to engage in sexual reproduction. Methodology/ Principle Findings In the present study, we investigated two hypotheses that can reconcile the apparent contradiction between the observed clonal population structure and the capacity to engage in sexual reproduction by analyzing the genome sequences of 123 T. cruzi isolates from a natural population in Arequipa, Peru. The distribution of polymorphic markers within and among isolates provides clear evidence of the occurrence of sexual reproduction. Large genetic segments are rearranged among chromosomes due to crossing over during meiosis leading to a decay in the genetic linkage among polymorphic markers compared to the expectations from a purely asexually-reproducing population. Nevertheless, the population structure appears clonal due to a high level of inbreeding during sexual reproduction which increases homozygosity, and thus reduces diversity, within each inbreeding lineage. Conclusions/ Significance These results effectively reconcile the apparent contradiction by demonstrating that the clonal population structure is derived not from infrequent sex in natural populations but from high levels of inbreeding. We discuss epidemiological consequences of this reproductive strategy on genome evolution, population structure, and phenotypic diversity of this medically important parasite., Author summary The rearrangement of alleles among individuals in a population during sexual reproduction maintains high allelic diversity within individuals in a population at polymorphic genes. Allelic diversity within individuals can be particularly important for parasites as it enhances their ability to evade host immune systems. Despite the potential benefits of sexual reproduction for parasites, natural populations of the protozoan parasite—and causative agent of human Chagas disease—Trypanosoma cruzi, exhibit clonal population structures indicative of asexual reproduction. This is particularly surprising as T. cruzi has the capacity for sexual reproduction. Here, we resolve this apparent contradiction by sequencing whole genomes of 123 T. cruzi isolates from a natural population in Arequipa, Peru. Evidence of past sexual reproduction and allelic rearrangements are common in this T. cruzi population. However, the majority of sexual reproduction events occur between close relatives resulting in an apparent clonal population structure. Sexual reproduction with distant relatives in areas with greater strain diversity has the potential to affect public health by increasing diversity in immune evasion genes within individuals and enhancing within-host survival, rapidly diversifying antigens that could affect the sensitivity of serological diagnostics, and by generating diversity in pathogenicity or drug resistance.

The vector of Chagas disease, Triatoma infestans, is largely controlled by the household application of pyrethroid insecticides. Because effective, large-scale insecticide application is costly and necessitates numerous trained personnel, alternative control techniques are badly needed. We compared the residual effect of organophosphate-based insecticidal paint (Inesfly 5A IGR™ (I5A)) to standard deltamethrin, and a negative control, against T. infestans in a simulated natural environment. We evaluated mortality, knockdown, and ability to take a blood meal among 5 th instar nymphs. I5A paint caused significantly greater mortality at time points up to nine months compared to deltamethrin (Fisher's Exact Test, p < 0.01 in all instances). A year following application, mortality among nymphs in the I5A was similar to those in the deltamethrin (χ2 = 0.76, df=1, p < 0.76). At months 0 and 1 after application, fewer nymphs exposed to deltamethrin took a blood meal compared to insects exposed to paint (Fisher's Exact Tests, p < 0.01 and p < 0.01, respectively). Insecticidal paint may provide an easily-applied means of protection against vectors of Chagas disease. Journal of Vector Ecology 38 (1): 6-11. 2013.

BACKGROUND: Chagas disease, a vector-borne disease transmitted by triatomine bugs and caused by the parasite Trypanosoma cruzi, affects millions of people in the Americas. In Arequipa, Peru, indoor residual insecticide spraying campaigns are routinely conducted to eliminate Triatoma infestans, the only vector in this area. Following insecticide spraying, there is risk of vector return and reinitiation of parasite transmission. Dogs are important reservoirs of T. cruzi and may play a role in reinitiating transmission in previously sprayed areas. Dogs may also serve as indicators of reemerging transmission. METHODS: We conducted a cross-sectional serological screening to detect T. cruzi antibodies in dogs, in conjunction with an entomological vector collection survey at the household level, in a disease endemic area that had been treated with insecticide 13 years prior. Spatial clustering of infected animals and vectors was assessed using Ripley's K statistic, and the odds of being seropositive for dogs proximate to infected colonies was estimated with multivariate logistic regression. RESULTS: There were 106 triatomine-infested houses (41.1%), and 45 houses infested with T. cruzi-infected triatomine insects (17.4%). Canine seroprevalence in the area was 12.3% (n=154); all seropositive dogs were 9 months old or older. We observed clustering of vectors carrying the parasite, but no clustering of seropositive dogs. The age- and sex-adjusted odds ratio between seropositivity to T. cruzi and proximity to an infected triatomine (

OBJETIVOS: Determinar la prevalencia de anticuerpos contra Trypanosoma cruzi en puérperas y la posible transmisión congénita de la enfermedad de Chagas en Arequipa, Perú, una zona donde esta enfermedad es endémica. MÉTODOS: Se estudió a las puérperas que dieron a luz entre diciembre de 2001 y julio de 2002 en tres hospitales (dos urbanos y uno rural) y cuatro centros de salud (tres rurales y uno urbano) del departamento de Arequipa, Perú. El estudio serológico comprendió el tamizaje de todas las puérperas para detectar anticuerpos contra T. cruzi mediante inmunofluorescencia indirecta (IFI); la prueba de inmunoadsorción enzimática (ELISA) y la titulación de anticuerpos IgG por IFI se usaron como pruebas confirmatorias. A las puérperas con seropositividad y a sus recién nacidos se les realizó la prueba de detección de anticuerpos IgM mediante IFI y se evaluó la presencia de infección mediante xenodiagnóstico (evaluada a los 30 y 60 días) y el micrométodo de Freilij. Los resultados se analizaron según la presencia del vector y de casos de enfermedad de Chagas en los lugares de nacimiento y de residencia de las puérperas. Dos neonatólogos evaluaron clínicamente a los recién nacidos para detectar anomalías y signos de enfermedad de Chagas congénita. RESULTADOS: La prevalencia general de enfermedad de Chagas en las 3 000 puérperas estudiadas fue de 0,73%; fue mayor en dos centros de salud ubicados en zonas rurales (2,2% en El Pedregal y 4,1% en La Joya) (P = 0,018) y la enfermedad estuvo asociada con el contacto directo previo con el vector (P < 0,05) y con el haber nacido en una zona considerada endémica (P < 0,01). Cuatro de las 20 puérperas con seropositividad (20%) tuvieron resultados positivos en el xenodiagnóstico. Ninguna conocía su estado de portadora de la infección y no se observaron síntomas o signos característicos de la enfermedad de Chagas aguda o crónica. En ninguna puérpera se detectaron anticuerpos IgM y solo un neonato (nacido de una madre sin parasitemia) presentó un título de IgM de 1/8, pero en los controles posteriores no se detectaron anticuerpos IgM o IgG. No se detectaron parásitos en la sangre de los neonatos por ninguno de los dos métodos empleados. De los 20 neonatos evaluados, uno tenía microcefalia y hepatoesplenomegalia y aunque tenía anticuerpos específicos IgG contra T. cruzi al nacer, estos desaparecieron a los dos meses; el crecimiento y el desarrollo de los demás recién nacidos fueron normales. CONCLUSIÓN: La prevalencia de enfermedad de Chagas en puérperas del departamento de Arequipa, Perú, es baja. No se encontraron casos de transmisión congénita intrauterina. Se recomienda diseñar estudios de detección prenatal que permitan evaluar a un mayor número de madres y en el que participen también las mujeres que dan a luz en sus domicilios.OBJECTIVES: To determine the prevalence of antibodies against Trypanosoma cruzi in puerperal women and to assess possible congenital transmission of Chagas' disease in the department of Arequipa, Peru, where the disease is endemic. METHODS: Women who had given birth between December 2001 and July 2002 in three hospitals (two urban and one rural) and four health centers (three rural and one urban) of the department of Arequipa, Peru, were studied. The serological study included screening all the puerperal women in order to detect antibodies against T. cruzi through indirect immunofluorescence (IIF), with confirmatory testing done with enzyme-linked immunosorbent assay (ELISA) testing and the titration of immunoglobulin G (IgG) antibodies by IIF. IIF tests to screen for immunoglobulin M (IgM) antibodies were done with the seropositive women and their newborns, and infection was evaluated through xenodiagnosis (evaluated at 30 and 60 days) and the direct micromethod of Freilij et al. The results were analyzed in terms of the presence of the vector and of cases of Chagas' disease in the places where the puerperal women had been born and where they were living. Two neonatologists clinically evaluated the newborns in order to detect abnormalities and signs of congenital Chagas' disease. RESULTS: The overall prevalence of Chagas' disease in the 3 000 puerperal women studied was 0.73%. Prevalence was highest in two health centers located in rural areas (2.2% in El Pedregal and 4.1% in La Joya) (P = 0.018). The disease was associated with previous direct contact with the vector (P < 0.05) and with having been born in an area considered endemic (P < 0.01). Four (20%) of the 20 seropositive puerperal women were also positive by xenodiagnosis. However, none of the women was aware of her infectious carrier state, and none showed the characteristic symptoms or signs of acute or chronic Chagas' disease. IgM antibodies were not detected in any of the puerperal women. One neonate (whose mother did not have evidence of parasitemia) presented an IgM titer of 1/8, but in later controls neither IgM nor IgG antibodies were detected. Parasites were not detected in the blood of the neonates by either of the two testing methods used. Of the 20 neonates evaluated, one presented microcephaly and hepatosplenomegaly; although the child had specific IgG antibodies against T. cruzi at birth, the antibodies were not present at the age of two months. The growth and development of the other 19 newborns were normal. CONCLUSIONS: The prevalence of Chagas' disease in puerperal women of the department of Arequipa, Peru, is low. No cases of intrauterine congenital transmission were found. We recommend carrying out studies on prenatal detection that evaluate more mothers and in which women who give birth at home also participate.

OBJETIVOS: Determinar la prevalencia de anticuerpos contra Trypanosoma cruzi en puerperas y la posible transmision congenita de la enfermedad de Chagas en Arequipa, Peru, una zona donde esta enfermedad es endemica. METODOS: Se estudio a las puerperas que dieron a luz entre diciembre de 2001 y julio de 2002 en tres hospitales (dos urbanos y uno rural) y cuatro centros de salud (tres rurales y uno urbano) del departamento de Arequipa, Peru. El estudio serologico comprendio el tamizaje de todas las puerperas para detectar anticuerpos contra T. cruzi mediante inmunofluorescencia indirecta (IFI); la prueba de inmunoadsorcion enzimatica (ELISA) y la titulacion de anticuerpos IgG por IFI se usaron como pruebas confirmatorias. A las puerperas con seropositividad y a sus recien nacidos se les realizo la prueba de deteccion de anticuerpos IgM mediante IFI y se evaluo la presencia de infeccion mediante xenodiagnostico (evaluada a los 30 y 60 dias) y el micrometodo de Freilij. Los resultados se analizaron segun la presencia del vector y de casos de enfermedad de Chagas en los lugares de nacimiento y de residencia de las puerperas. Dos neonatologos evaluaron clinicamente a los recien nacidos para detectar anomalias y signos de enfermedad de Chagas congenita. RESULTADOS: La prevalencia general de enfermedad de Chagas en las 3 000 puerperas estudiadas fue de 0,73%; fue mayor en dos centros de salud ubicados en zonas rurales (2,2% en El Pedregal y 4,1% en La Joya) (P = 0,018) y la enfermedad estuvo asociada con el contacto directo previo con el vector (P < 0,05) y con el haber nacido en una zona considerada endemica (P < 0,01). Cuatro de las 20 puerperas con seropositividad (20%) tuvieron resultados positivos en el xenodiagnostico. Ninguna conocia su estado de portadora de la infeccion y no se observaron sintomas o signos caracteristicos de la enfermedad de Chagas aguda o cronica. En ninguna puerpera se detectaron anticuerpos IgM y solo un neonato (nacido de una madre sin parasitemia) presento un titulo de IgM de 1/8, pero en los controles posteriores no se detectaron anticuerpos IgM o IgG. No se detectaron parasitos en la sangre de los neonatos por ninguno de los dos metodos empleados. De los 20 neonatos evaluados, uno tenia microcefalia y hepatoesplenomegalia y aunque tenia anticuerpos especificos IgG contra T. cruzi al nacer, estos desaparecieron a los dos meses; el crecimiento y el desarrollo de los demas recien nacidos fueron normales. CONCLUSION: La prevalencia de enfermedad de Chagas en puerperas del departamento de Arequipa, Peru, es baja. No se encontraron casos de transmision congenita intrauterina. Se recomienda disenar estudios de deteccion prenatal que permitan evaluar a un mayor numero de madres y en el que participen tambien las mujeres que dan a luz en sus domicilios.

Background Vector-borne transmission of Trypanosoma cruzi is seen exclusively in the Americas where an estimated 8 million people are infected with the parasite. Significant research in southern Peru has been conducted to understand T. cruzi infection and vector control, however, much less is known about the burden of infection and epidemiology in northern Peru. Methodology A cross-sectional study was conducted to estimate the seroprevalence of T. cruzi infection in humans (n=611) and domestic animals [dogs (n=106) and guinea pigs (n=206)] in communities of Cutervo Province, Peru. Sampling and diagnostic strategies differed according to species. An entomological household study (n=208) was conducted to identify the triatomine burden and species composition, as well as the prevalence of T. cruzi in vectors. Electrocardiograms (EKG) were performed on a subset of participants (n=90 T. cruzi infected participants and 170 age and sex-matched controls). The seroprevalence of T. cruzi among humans, dogs, and guinea pigs was 14.9% (95% CI: 12.2 – 18.0%), 19.8% (95% CI: 12.7- 28.7%) and 3.3% (95% CI: 1.4 – 6.9%) respectively. In one community, the prevalence of T. cruzi infection was 17.2% (95% CI: 9.6 - 24.7%) among participants < 15 years, suggesting recent transmission. Increasing age, positive triatomines in a participant's house, and ownership of a T. cruzi positive guinea pig were independent correlates of T. cruzi infection. Only one species of triatomine was found, Panstrongylus lignarius, formerly P. herreri. Approximately forty percent (39.9%, 95% CI: 33.2 - 46.9%) of surveyed households were infested with this vector and 14.9% (95% CI: 10.4 - 20.5%) had at least one triatomine positive for T. cruzi. The cardiac abnormality of right bundle branch block was rare, but only identified in seropositive individuals. Conclusions Our research documents a substantial prevalence of T. cruzi infection in Cutervo and highlights a need for greater attention and vector control efforts in northern Peru., Author Summary Chagas disease causes significant morbidity and mortality throughout Central and South America. The epidemiology and control of this disease is subject to unique regional particularities, including the behavior and ecology of the local insect vector species. Significant resources have been allocated towards research and control efforts in southern Peru, yet very little is known about the prevalence and epidemiology of Trypanosoma cruzi in northern Peru. Our study highlights significant T. cruzi infection in northern Peru and is one of the first to document substantial transmission by the insect Panstrongylus lignarius. Our results illustrate major gaps in knowledge and the need for public health interventions targeted at Chagas disease in the region of Cutervo Province of northern Peru.

The increasing rate of biological invasions resulting from human transport or human-mediated changes to the environment has had devastating ecological and public health consequences. The kissing bug, Triatoma infestans, has dispersed through the Peruvian city of Arequipa. The biological invasion of this insect has resulted in a public health crisis, putting thousands of residents of this city at risk of infection by Trypanosoma cruzi and subsequent development of Chagas disease. Here, we show that populations of Tria. infestans in geographically distinct districts within and around this urban centre share a common recent evolutionary history although current gene flow is restricted even between proximal sites. The population structure among the Tria. infestans in different districts is not correlated with the geographical distance between districts. These data suggest that migration among the districts is mediated by factors beyond the short-range migratory capabilities of Tria. infestans and that human movement has played a significant role in the structuring of the Tria. infestans population in the region. Rapid urbanization across southern South America will continue to create suitable environments for Tria. infestans, and knowledge of its urban dispersal patterns may play a fundamental role in mitigating human disease risk.

Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.- Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.- restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.

Statistical methods such as latent class analysis can estimate the sensitivity and specificity of diagnostic tests when no perfect reference test exists. Traditional latent class methods assume a constant disease prevalence in one or more tested populations. When the risk of disease varies in a known way, these models fail to take advantage of additional information that can be obtained by measuring risk factors at the level of the individual. We show that by incorporating complex field-based epidemiologic data, in which the disease prevalence varies as a continuous function of individual-level covariates, our model produces more accurate sensitivity and specificity estimates than previous methods. We apply this technique to several simulated populations and to actual Chagas disease test data from a community near Arequipa, Peru. Results from our model estimate that the first-line enzyme-linked immunosorbent assay has a sensitivity of 78% (95% CI: 62-100%) and a specificity of 100% (95% CI: 99-100%). The confirmatory immunofluorescence assay is estimated to be 73% sensitive (95% CI: 65-81%) and 99% specific (95% CI: 96-100%).

We used genus/species specific PCRs to determine the temporal persistence of host DNA in Triatoma infestans experimentally fed on blood from six common vertebrate species: humans, domestic dogs, guinea pigs, chickens, mice, and pigs. Twenty third or fourth instar nymphs per animal group were allowed to feed to engorgement, followed by fasting-maintenance in the insectary. At 7, 14, 21, or 28 days post-feeding, the midgut contents from five triatomines per group were tested with the respective PCR assay. DNA from all vertebrate species was detected in at least four of five study nymphs at seven and 14 days post-feeding. DNA of humans, domestic dogs, guinea pigs, pigs, and chickens were more successfully detected (80-100%) through day 21, and less successfully (20-100%) at day 28. Findings demonstrate that species-specific PCRs can consistently identify feeding sources of T. infestans within two weeks, a biologically relevant time interval. Se utilizó pruebas PCR género o especie específicas para determinar la persistencia temporal de ADN del hospedero en el contenido intestinal de Triatoma infestans que fueron alimentados experimentalmente con sangre de seis vertebrados muy frecuentemente asociados a enfermedad de Chagas: humano, perro, cobayo, pollo, ratón, y cerdo. Se emplearon 20 ninfas de tercer y cuarto estadio por cada especie de hospedero. Fueron alimentados a saciedad y mantenidas en el insectario sin alimentación posterior. Se obtuvo el contenido intestinal de cinco triatominos por cada grupo a los 7, 14, 21 y 28 días post - alimentación, que fueron evaluados con los respectivos PCRs específicos. El ADN de todos los vertebrados fue detectado en al menos 4 de 5 ninfas evaluadas a los 7 y 14 días post - alimentación. El ADN de humano, perro, cobayo, cerdo y pollo fue detectado exitosamente (80-100%) hasta el día 21 y con menos éxito (20-100%) en el día 28. Estos resultados demuestran que PCRs específicos para cada especie de hospedero pueden identificar consistentemente la fuente de alimentación de T. infestans dentro de las dos semanas post - alimentación, siendo un intervalo de tiempo biológicamente relevante.

Insecticide-impregnated nets can kill triatomine bugs, but it remains unclear whether they can protect against Chagas disease transmission. In a field trial in Quequeña, Peru, sentinel guinea pigs placed in intervention enclosures covered by deltamethrin-treated nets showed significantly lower antibody responses to saliva of Triatoma infestans compared with animals placed in pre-existing control enclosures. Our results strongly suggest that insecticide-treated nets prevent triatomine bites and can thereby protect against infection with Trypanosoma cruzi. Anti-salivary immunoassays are powerful new tools to evaluate intervention strategies against Chagas disease.

Background The history of Chagas disease control in Peru and many other nations is marked by scattered and poorly documented vector control campaigns. The complexities of human migration and sporadic control campaigns complicate evaluation of the burden of Chagas disease and dynamics of Trypanosoma cruzi transmission. Methodology/Principal Findings We conducted a cross-sectional serological and entomological study to evaluate temporal and spatial patterns of T. cruzi transmission in a peri-rural region of La Joya, Peru. We use a multivariate catalytic model and Bayesian methods to estimate incidence of infection over time and thereby elucidate the complex history of transmission in the area. Of 1,333 study participants, 101 (7.6%; 95% CI: 6.2–9.0%) were confirmed T. cruzi seropositive. Spatial clustering of parasitic infection was found in vector insects, but not in human cases. Expanded catalytic models suggest that transmission was interrupted in the study area in 1996 (95% credible interval: 1991–2000), with a resultant decline in the average annual incidence of infection from 0.9% (95% credible interval: 0.6–1.3%) to 0.1% (95% credible interval: 0.005–0.3%). Through a search of archival newspaper reports, we uncovered documentation of a 1995 vector control campaign, and thereby independently validated the model estimates. Conclusions/Significance High levels of T. cruzi transmission had been ongoing in peri-rural La Joya prior to interruption of parasite transmission through a little-documented vector control campaign in 1995. Despite the efficacy of the 1995 control campaign, T. cruzi was rapidly reemerging in vector populations in La Joya, emphasizing the need for continuing surveillance and control at the rural-urban interface., Author Summary The historically rural problem of Chagas disease is increasing in urban areas in Latin America. Peri-rural development may play a critical role in the urbanization of Chagas disease and other parasitic infections. We conducted a cross-sectional study in an urbanizing rural area in southern Peru, and we encountered a complex history of Chagas disease in this peri-rural environment. Specifically, we discovered: (1) long-standing parasite transmission leading to substantial burden of infection; (2) interruption in parasite transmission resulting from an undocumented insecticide application campaign; (3) relatively rapid re-emergence of parasite-infected vector insects resulting from an unsustained control campaign; (4) extensive migration among peri-rural inhabitants. Long-standing parasite infection in peri-rural areas with highly mobile populations provides a plausible mechanism for the expansion of parasite transmission to nearby urban centers. Lack of commitment to control campaigns in peri-rural areas may have unforeseen and undesired consequences for nearby urban centers. Novel methods and perspectives are needed to address the complexities of human migration and erratic interventions.

Control of the Chagas disease vector, Triatoma infestans, relies on the application of pyrethroid insecticides, especially deltamethrin. We performed laboratory studies to determine whether a T. infestans nymph that comes into contact with a deltamethrin-treated surface horizontally transfers the insecticide to subsequent triatomines. We found that a triatomine that walks on a deltamethrin-treated surface for a short period of time has the ability to transport the insecticide in concentrations sufficient to kill other triatomines with which it comes into contact. The effect was limited to high-density environments, and mortality as a result of secondary exposure was greater among second-instar nymphs compared with fifth-instar nymphs. Our results suggest that deltamethrin could be killing triatomines through both direct and indirect contact, although it remains unclear whether the phenomenon occurs in natural conditions.

Introducción: Existe escasa información y no actualizada sobre la presencia de triatominos en el departamento de San Martín, zona amazónica de nuestro país, y es importante actualizarla, ya que hay reportes recientes de casos de enfermedad de Chagas procedentes de la amazonía peruana cuyos vectores serían triatominos silvestres. Objetivos: Conocer la distribución geográfica de triatominos hematófagos en Huallaga y Picota, San Martín, Perú. Determinar los índices entomológicos, sus hábitos alimenticios y caracterizar los tripanosomatídeos que portan. Diseño: Investigación descriptiva observacional. Lugar: Red de Salud Huallaga y Picota, Laboratorio Regional de la Región de Salud San Martín. Laboratorios del Instituto Nacional de Salud. Participantes: Viviendas de 6 distritos de las provincia de Huallaga y 9 de Picota, del Departamento de San Martín. Intervenciones: La búsqueda de triatominos fue en 3 362 viviendas de áreas periurbanas y rurales. Mediante la prueba de precipitina, se determinó la fuente de alimentación. Se aisló tripanosomatídeos mediante infección en ratones Balb/C e inoculación en medio agar sangre, y por PCR se caracterizó el genoma por amplificación del espaciador intergénico del gen miniexón. Principales medidas de resultados: Determinar si Huallaga y Picota, San Martín, Perú, son zonas de riesgo de enfermedad de Chagas. Resultados: En 46 viviendas, de 3 362 muestreadas se capturó 53 triatominos adultos pertenecientes a Rhodnius pictipes (56,6%), Panstrongylus geniculatus (41,5%) y Eratyrus mucronatus (1,9%). Se reporta P. geniculatus por primera vez para las dos provincias. Los índices de infestación para ambas provincias fueron 1,4% y 3,8%, respectivamente, y corresponden al índice de infección natural; mientras que el índice de dispersión fue 30,1%. En R. pictipes se identificó cinco fuentes de alimentación (pollo, humano, perro, cobayo y gato) y en P. geniculatus un perfil alimentario mixto: perro/gato/pollo. Los Trypanosoma spp. aislados de R. pictipes, procedente de Sacanche (Huallaga), corresponden a TCI (350 pb); resultado similar se obtuvo en los 10 clones de este aislamiento. Conclusiones: Los resultados nos indican que, en las localidades estudiadas, R. pictipes y P. geniculatus estarían pasando por un proceso de domiciliación y que en las localidades de Huallaga R. pictipes sería el principal vector de Trypanosoma cruzi TC1; por lo que, dichas localidades estarían consideradas como zonas de riesgo de enfermedad de Chagas. Introduction: There is little and not recent information on the presence of triatomine bugs in San Martin department, Amazon area of our country where Chagas disease has been reported by wild triatomine insects as vectors. Objectives: To determine the geographic distribution of hematophagous triatomine insects in Huallaga and Picota provinces, San Martin department, Peru. To determine the domiciliary infestation by triatomine vectors indexes, trypano-triatomine infection as well as their dispersion. To determine the insects feeding source and to genetically characterize the tripanosomatidae they carry. Design: Observational and descriptive study. Setting: Health and Laboratories region networks. National Institute of Health laboratories. Participants: Houses from 6 districts of Huallaga province and 9 from Picota province (San Martin department). Interventions: Triatomine insect’s collection was carried out in 3 362 homes located in semi-urban and rural zones accounting for 30% of the total area of studied provinces. Feeding preferences were assessed by a precipitin test using specific antibodies against guinea pig, dog, cat, chicken and human beings sera. Trypanosomes were isolated by inoculation of stool samples from infected insects in Balb/c mice and using axenic culture media. Genetic characterization was done by PCR amplification of the genome by gene intergenic spacer of the miniexon. Main outcome measures: To assess if Huallaga and Picota, San Martin, Peru, are Chagas disease risk areas. Results: In 46 of the 3 368 homes visited, 53 triatomine adults were collected belonging to the species Rhodnius pictipes (56,6%), Panstrongylus geniculatus (41,5%) and Eratyrus mucronatus (1,9%). For the first time, P. geniculatus was reported in both provinces included in the study. Domiciliary infestation index was 1,4%, trypano-triatomine infection index was 3,8% and dispersion index was 30,1%. For R. pictipes 5 sources of food were identified (chicken, human, dog, guinea pig and cat) and for P. geniculatus, a mix feed: dog, cat and chicken. Trypanosoma spp. isolates of R. pictipes from Sacanche (Huallaga) belonged to T. cruzi TCI (350 pb); similar result was obtained in 10 clones studied. Conclusions: Results indicate that R. pictipes and P. geniculatus are undergoing colonization of the regions included in this study. Within the province of Huallaga, R. pictipes is the predominant vector of T. cruzi; therefore these locations must be considered high risk areas for Chagas disease.

Faeces-mediated transmission of Trypanosoma cruzi (the aetiological agent of Chagas disease) by triatomine insects is extremely inefficient. Still, the parasite emerges frequently, and has infected millions of people and domestic animals. We synthesize here the results of field and laboratory studies of T. cruzi transmission conducted in and around Arequipa, Peru. We document the repeated occurrence of large colonies of triatomine bugs (more than 1000) with very high infection prevalence (more than 85%). By inoculating guinea pigs, an important reservoir of T. cruzi in Peru, and feeding triatomine bugs on them weekly, we demonstrate that, while most animals quickly control parasitaemia, a subset of animals remains highly infectious to vectors for many months. However, we argue that the presence of these persistently infectious hosts is insufficient to explain the observed prevalence of T. cruzi in vector colonies. We posit that seasonal rains, leading to a fluctuation in the price of guinea pig food (alfalfa), leading to annual guinea pig roasts, leading to a concentration of vectors on a small subpopulation of animals maintained for reproduction, can propel T. cruzi through vector colonies and create a considerable force of infection for a pathogen whose transmission might otherwise fizzle out.

To determine the prevalence of antibodies against Trypanosoma cruzi in puerperal women and to assess possible congenital transmission of Chagas' disease in the department of Arequipa, Peru, where the disease is endemic.Women who had given birth between December 2001 and July 2002 in three hospitals (two urban and one rural) and four health centers (three rural and one urban) of the department of Arequipa, Peru, were studied. The serological study included screening all the puerperal women in order to detect antibodies against T. cruzi through indirect immunofluorescence (IIF), with confirmatory testing done with enzyme-linked immunosorbent assay (ELISA) testing and the titration of immunoglobulin G (IgG) antibodies by IIF. IIF tests to screen for immunoglobulin M (IgM) antibodies were done with the seropositive women and their newborns, and infection was evaluated through xenodiagnosis (evaluated at 30 and 60 days) and the direct micromethod of Freilij et al. The results were analyzed in terms of the presence of the vector and of cases of Chagas' disease in the places where the puerperal women had been born and where they were living. Two neonatologists clinically evaluated the newborns in order to detect abnormalities and signs of congenital Chagas' disease.The overall prevalence of Chagas' disease in the 3 000 puerperal women studied was 0.73%. Prevalence was highest in two health centers located in rural areas (2.2% in El Pedregal and 4.1% in La Joya) (P=0.018). The disease was associated with previous direct contact with the vector (P0.05) and with having been born in an area considered endemic (P0.01). Four (20%) of the 20 seropositive puerperal women were also positive by xenodiagnosis. However, none of the women was aware of her infectious carrier state, and none showed the characteristic symptoms or signs of acute or chronic Chagas' disease. IgM antibodies were not detected in any of the puerperal women. One neonate (whose mother did not have evidence of parasitemia) presented an IgM titer of 1/8, but in later controls neither IgM nor IgG antibodies were detected. Parasites were not detected in the blood of the neonates by either of the two testing methods used. Of the 20 neonates evaluated, one presented microcephaly and hepatosplenomegaly; although the child had specific IgG antibodies against T. cruzi at birth, the antibodies were not present at the age of two months. The growth and development of the other 19 newborns were normal.The prevalence of Chagas' disease in puerperal women of the department of Arequipa, Peru, is low. No cases of intrauterine congenital transmission were found. We recommend carrying out studies on prenatal detection that evaluate more mothers and in which women who give birth at home also participate.

Modern cities represent one of the fastest growing ecosystems on the planet. Urbanization occurs in stages; each stage characterized by a distinct habitat that may be more or less susceptible to the establishment of disease vector populations and the transmission of vector-borne pathogens. We performed longitudinal entomological and epidemiological surveys in households along a 1900 × 125 m transect of Arequipa, Peru, a major city of nearly one million inhabitants, in which the transmission of Trypanosoma cruzi , the aetiological agent of Chagas disease, by the insect vector Triatoma infestans , is an ongoing problem. The transect spans a cline of urban development from established communities to land invasions. We find that the vector is tracking the development of the city, and the parasite, in turn, is tracking the dispersal of the vector. New urbanizations are free of vector infestation for decades. T. cruzi transmission is very recent and concentrated in more established communities. The increase in land tenure security during the course of urbanization, if not accompanied by reasonable and enforceable zoning codes, initiates an influx of construction materials, people and animals that creates fertile conditions for epidemics of some vector-borne diseases.

With increasing urbanization vector-borne diseases are quickly developing in cities, and urban control strategies are needed. If streets are shown to be barriers to disease vectors, city blocks could be used as a convenient and relevant spatial unit of study and control. Unfortunately, existing spatial analysis tools do not allow for assessment of the impact of an urban grid on the presence of disease agents. Here, we first propose a method to test for the significance of the impact of streets on vector infestation based on a decomposition of Moran's spatial autocorrelation index; and second, develop a Gaussian Field Latent Class model to finely describe the effect of streets while controlling for cofactors and imperfect detection of vectors. We apply these methods to cross-sectional data of infestation by the Chagas disease vector Triatoma infestans in the city of Arequipa, Peru. Our Moran's decomposition test reveals that the distribution of T. infestans in this urban environment is significantly constrained by streets (p, Author Summary Chagas disease is a major parasitic disease in Latin America. It is transmitted by Triatoma infestans an insect common in Arequipa, the second largest city in Peru. We propose a method to demonstrate that streets strongly affect the spatial distribution of infestation by this insect in Arequipa. The effect of streets may be due to several external factors: 1) houses on different sides of a street may not be equally welcoming to the insects due to the presence of certain materials or animals, 2) people inspecting houses on the two sides of a street may not be equally efficient, and, 3) insects may disperse to neighboring houses but rarely reach houses across a street. We take these aspects into account in a second analysis and confirm that streets are important barriers to these insects. Our finding should allow for improvements in the control of insects that transmit Chagas disease in cities. More generally, our methods can be applied to other pests and disease vectors to better understand and control epidemics in cities.

Background Chagas disease is endemic in the rural areas of southern Peru and a growing urban problem in the regional capital of Arequipa, population ∼860,000. It is unclear how to implement cost-effective screening programs across a large urban and periurban environment. Methods We compared four alternative screening strategies in 18 periurban communities, testing individuals in houses with 1) infected vectors; 2) high vector densities; 3) low vector densities; and 4) no vectors. Vector data were obtained from routine Ministry of Health insecticide application campaigns. We performed ring case detection (radius of 15 m) around seropositive individuals, and collected data on costs of implementation for each strategy. Results Infection was detected in 21 of 923 (2.28%) participants. Cases had lived more time on average in rural places than non-cases (7.20 years versus 3.31 years, respectively). Significant risk factors on univariate logistic regression for infection were age (OR 1.02; p = 0.041), time lived in a rural location (OR 1.04; p = 0.022), and time lived in an infested area (OR 1.04; p = 0.008). No multivariate model with these variables fit the data better than a simple model including only the time lived in an area with triatomine bugs. There was no significant difference in prevalence across the screening strategies; however a self-assessment of disease risk may have biased participation, inflating prevalence among residents of houses where no infestation was detected. Testing houses with infected-vectors was least expensive. Ring case detection yielded four secondary cases in only one community, possibly due to vector-borne transmission in this community, apparently absent in the others. Conclusions Targeted screening for urban Chagas disease is promising in areas with ongoing vector-borne transmission; however, these pockets of epidemic transmission remain difficult to detect a priori. The flexibility to adapt to the epidemiology that emerges during screening is key to an efficient case detection intervention. In heterogeneous urban environments, self-assessments of risk and simple residence history questionnaires may be useful to identify those at highest risk for Chagas disease to guide diagnostic efforts., Author Summary In the wake of emerging T. cruzi infection in children of periurban Arequipa, Peru, we conducted a prospective field trial to evaluate alternative targeted screening strategies for Chagas disease across the city. Using insect vector data that is routinely collected during Ministry of Health insecticide application campaigns in 3 periurban districts of Arequipa, we separated into 4 categories those households with 1) infected vectors; 2) high vector densities; 3) low vector densities; and 4) no vectors. Residents of all infected-vector households and a random sample of those in the other 3 categories were invited for serological screening for T. cruzi infection. Subsequently, all residents of households within a 15-meter radius of detected seropositive individuals were invited to be screened in a ring case-detection scheme. Of 923 participants, 21 (2.28%) were seropositive. There were no significant differences in prevalence across the 4 screening strategies, indicating that household entomologic factors alone could not predict the risk of infection. Indeed, the most predictive variable of infection was the number of years a person lived in a location with triatomine insects. Therefore, a simple residence history questionnaire may be a useful screening tool in large, diverse urban environments with emerging Chagas disease.