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RESUMEN Objetivos. Evaluar la variación de los perfiles hematológicos antes, durante y después del tratamiento de pacientes infectados con malaria no complicada por Plasmodium vivax (Pv) y P. falciparum (Pf) en una población de la región Loreto. Materiales y métodos. El estudio se realizó entre 2010 y 2012, en Zungarococha (Iquitos). Los 425 participantes tuvieron tres visitas (visita 1-día 0-antes del tratamiento, visita 2-día 7-durante tratamiento, visita 3-día 28-después del tratamiento), hemograma completo, diagnóstico microscópico y molecular (PCR). Resultados. En la primera visita, se encontraron 93 (21,9%) positivos a Pv y 34 (8,0%) a Pf. Todos los positivos mostraron una reducción en los indicadores hematológicos de hematocrito, recuento de glóbulos blancos (RGB), neutrófilos abastonados y segmentados, eosinófilos y plaquetas (p

To evaluate the variation of hematological profiles of patients infected with uncomplicated Plasmodium vivax (Pv) and P. falciparum (Pf) malaria before, during and after treatment in a population of the Loreto region.This study was conducted between 2010 and 2012, in Zungarococha (Iquitos). The 425 participants had three visits (visit 1-day 0-before treatment, visit 2-day 7-during treatment, visit 3-day 28-after treatment), complete blood count, microscopic and molecular diagnosis (PCR).At the first visit, 93 (21.9%) participants were found positive for Pv and 34 (8.0%) for Pf. All positives showed a reduction in hematocrit, white blood cell count (WBC), ablated and segmented neutrophils, eosinophils and platelets (p0.001) compared to the negative group. A higher percentage of ablated neutrophils was found in Pf and segmented neutrophils in Pv compared to the negative group. Variations in hematological profiles were observed after treatment for both species; ablated neutrophils decreased, platelets increased, eosinophils increased at day 7 and declined at day 28, hematocrit and segmented neutrophils decreased at day 7 and normalized at day 28. Interspecies differences over time showed a bigger daily decrease in ablated neutrophils in Pv-infected when compared to Pf.The hematological profile in uncomplicated malaria-positive patients varies over time during and after treatment. These are indicators of disease progression and help in the therapeutic surveillance of Plasmodium-infected patients.Evaluar la variación de los perfiles hematológicos antes, durante y después del tratamiento de pacientes infectados con malaria no complicada por Plasmodium vivax (Pv) y P. falciparum (Pf) en una población de la región Loreto.El estudio se realizó entre 2010 y 2012, en Zungarococha (Iquitos). Los 425 participantes tuvieron tres visitas (visita 1-día 0-antes del tratamiento, visita 2-día 7-durante tratamiento, visita 3-día 28-después del tratamiento), hemograma completo, diagnóstico microscópico y molecular (PCR).En la primera visita, se encontraron 93 (21,9%) positivos a Pv y 34 (8,0%) a Pf. Todos los positivos mostraron una reducción en los indicadores hematológicos de hematocrito, recuento de glóbulos blancos (RGB), neutrófilos abastonados y segmentados, eosinófilos y plaquetas (p0.001) en comparación con el grupo negativo. Se encontró un porcentaje mayor de neutrófilos abastonados en Pf y de neutrófilos segmentados en Pv comparado al grupo negativo. Se observó variaciones en los perfiles hematológicos después del tratamiento para ambas especies, los neutrófilos abastonados disminuyeron, las plaquetas aumentaron, los eosinófilos se incrementaron al día 7 y decaen el día 28, el hematocrito y los neutrófilos segmentados disminuyeron al día 7 y se normalizaron el día 28. Las diferencias entre especies en el tiempo mostraron una disminución diaria de neutrófilos abastonados en infectados con Pv que en Pf.El perfil hematológico en pacientes positivos a malaria no complicada varía en el tiempo durante y después del tratamiento. Estos son indicadores de la progresión de la enfermedad y ayudan en la vigilancia terapéutica de pacientes infectados con Plasmodium.

Background Diagnosis of toxoplasmic encephalitis (TE) is challenging under the best clinical circumstances. The poor clinical sensitivity of quantitative polymerase chain reaction (qPCR) for Toxoplasma in blood and CSF and the limited availability of molecular diagnostics and imaging technology leaves clinicians in resource-limited settings with few options other than empiric treatment. Methology/principle findings Here we describe proof of concept for a novel urine diagnostics for TE using Poly-N-Isopropylacrylamide nanoparticles dyed with Reactive Blue-221 to concentrate antigens, substantially increasing the limit of detection. After nanoparticle-concentration, a standard western blotting technique with a monoclonal antibody was used for antigen detection. Limit of detection was 7.8pg/ml and 31.3pg/ml of T. gondii antigens GRA1 and SAG1, respectively. To characterize this diagnostic approach, 164 hospitalized HIV-infected patients with neurological symptoms compatible with TE were tested for 1) T. gondii serology (121/147, positive samples/total samples tested), 2) qPCR in cerebrospinal fluid (11/41), 3) qPCR in blood (10/112), and 4) urinary GRA1 (30/164) and SAG1 (12/164). GRA1 appears to be superior to SAG1 for detection of TE antigens in urine. Fifty-one HIV-infected, T. gondii seropositive but asymptomatic persons all tested negative by nanoparticle western blot and blood qPCR, suggesting the test has good specificity for TE for both GRA1 and SAG1. In a subgroup of 44 patients, urine samples were assayed with mass spectrometry parallel-reaction-monitoring (PRM) for the presence of T. gondii antigens. PRM identified antigens in 8 samples, 6 of which were concordant with the urine diagnostic. Conclusion/significances Our results demonstrate nanoparticle technology’s potential for a noninvasive diagnostic test for TE. Moving forward, GRA1 is a promising target for antigen based diagnostics for TE., Author summary Toxoplasmic Encephalitis is a debilitating, yet highly treatable illness, classically seen in person living with HIV lacking treatment. Prompt diagnosis ensures the best outcome possible for patients, but remains a challenge: requiring invasive specimen collection, lacking necessary clinical sensitivity, demanding significant technical skills, and substantial infrastructure. Here we offer proof of concept of a diagnostic approach that is minimally invasive, using a urine-based approach that concentrates T. gondii antigens with hydrogel mesh nanoparticles to improve analytical sensitivity for detection by western blot.

Background Malaria in Peru is concentrated in the Amazon region, especially in Loreto, and transmission is focused in rural and peri-urban communities. The government has approved a malaria elimination plan with a community approach and seeks to reduce the risk of transmission through preventive interventions, but asymptomatic and low-parasite-density infections are challenges for disease control and elimination. IgG antibodies play a critical role in combating infection through their ability to reduce parasitaemia and clinical symptoms. In particular, IgG subclasses have important roles in controlling malaria disease and may provide new insight into the development of malaria control strategies and understanding of malaria transmission. Through the use of excreted-secreted antigens from Plasmodium falciparum, were evaluated the responses of the four IgG subclasses in symptomatic and asymptomatic malarial infections. Results Higher levels of whole IgG were observed in asymptomatic carriers (P

The efficacy of tests with L. ( L .) infantum excreted-secreted antigens (ESA) to detect canine leishmaniasis (CanL) was evaluated using immunoblotting (ESA-blot), ELISA (ESA-ELISA) and ELISA with alkaline extract from promastigotes (PAE). Of one hundred fifty-five domestic dogs tested, 100 were suspected of CanL, 23 had other diseases and 32 were healthy. Sera from the dogs suspected of CanL were tested by immunohistochemistry (IHC), and 54% were confirmed to be infected by L. ( L. ) infantum (38 symptomatic and 16 asymptomatic). Of these, 100% were positive by ESA-blot, ESA-ELISA and PAE-ELISA. In the ESA-blot their sera recognized polypeptides in the 26.5–31.5 kDa region. Of the 46% of dogs with negative IHC, 44–53% tested positive in all three tests irrespective of clinical status. The twenty-three dogs with other diseases were negative by ESA-blot, but sera from 9% and 26% of them reacted with ESA-ELISA and PAE-ELISA, respectively. The 32 healthy dogs were negative in all the tests. ESA-blot showed good correlation with IHC in the detection of CanL and a high specificity index.

The analysis of promastigote excreted–secreted antigen (ESA) reactivity with 53 visceral leishmaniasis (VL) cases showed that each sample reacted regardless of the antigen or the Leishmania species used in enzyme-linked immunosorbent assay (ELISA) displayed 100% positivity with the L. (L.) chagasi ESA-blot recognizing bands of molecular weight ranging from 26.5 to 31.5 kDa. The analysis of 160 non-visceral cases showed that 5% of the samples cross-reacted with the L. (L.) chagasi ESA-ELISA and 9.4% reacted with the ESA isolated from L. (L.) amazonensis and L. (V.) braziliensis, whereas a high cross-reaction ranging from 24.4% to 25% was observed with total crude promastigote antigens (PRO-ELISA). The ESA-blot of L. (L.) chagasi tested with non-visceral sera samples showed a cross-reaction with 8.8% of cases; most of these cases represented tegumentary leishmaniasis and only one acute chagasic case. These data lead us to recommend the use of ESA as an alternative antigen in VL diagnosis.

Molecular tools to distinguish strains of Plasmodium vivax are important for studying the epidemiology of malaria transmission. Two sets of markers-tandem repeat (TR) polymorphisms and MSP3α-were used to study Plasmodium vivax in patients in the Peruvian Amazon region of Iquitos. Of 110 patients, 90 distinct haplotypes were distinguished using 9 TR markers. An MSP3α polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using HhaI and AluI revealed 8 and 9 profiles, respectively, and 36 profiles when analyzed in combination. Combining TR and PCR-RFLP markers, 101 distinct molecular profiles were distinguished among these 110 patients. Nine TR markers arrayed along a 100 kB stretch of a P. vivax chromosome containing the gene for circumsporozoite protein showed non-linear linkage disequilibrium (I(SA) = 0.03, P = 0.001). These findings demonstrate the potential use of TR markers for molecular epidemiology studies.

Chagas disease affects 8-11 million people throughout the Americas. Early detection is crucial for timely treatment and to prevent non-vectorial transmission. Recombinant antigen-based rapid tests had high sensitivity and specificity in laboratory evaluations, but no Peruvian specimens were included in previous studies. We evaluated Stat-Pak and Trypanosoma Detect rapid tests in specimens from Bolivia and Peru. Specimens positive by three conventional assays were confirmed positives; specimens negative by two or more assays were confirmed negatives. In Bolivian specimens, Stat-Pak and Trypanosoma Detect tests were 87.5% and 90.7% sensitive, respectively; both showed 100% specificity. Sensitivity in Peruvian specimens was much lower: 26.6-33.0% (Stat-Pak) and 54.3-55.2% (Trypanosoma Detect); both had specificities > 98%. Even in Bolivian specimens, these sensitivities are inadequate for stand-alone screening. The low sensitivity in Peru may be related to parasite strain differences. Chagas disease rapid tests should be field tested in each geographic site before widespread implementation for screening. Copyright

Anopheles darlingi is the most important malaria vector in the Amazon basin of South America, and is capable of transmitting both Plasmodium falciparum and P. vivax. To understand the genetic structure of this vector in the Amazonian region of Peru, a simple polymerase chain reaction (PCR)-based test to identify this species of mosquito was used. A random amplified polymorphic DNA-PCR was used to study genetic variation at the micro-geographic level in nine geographically separate populations of An. darlingi collected in areas with different degrees of deforestation surrounding the city of Iquitos. Within-population genetic diversity in nine populations, as quantified by the expected heterozygosity (HE), ranged from 0.27 to 0.32. Average genetic distance (FST) among these populations was 0.017. These results show that the nine studied populations are highly homogeneous, suggesting that strategies can be developed to combat this malaria vector as a single epidemiologic unit.

To examine the impact of tropical rain-forest destruction on malaria, we conducted a year-long study of the rates at which the primary malaria vector in the Amazon, Anopheles darlingi, fed on humans in areas with varying degrees of ecological alteration in the Peruvian Amazon. Mosquitoes were collected by human biting catches along the Iquitos-Nauta road at sites selected for type of vegetation and controlled for human presence. Deforested sites had an A. darlingi biting rate that was more than 278 times higher than the rate determined for areas that were predominantly forested. Our results indicate that A. darlingi displays significantly increased human-biting activity in areas that have undergone deforestation and development associated with road development.

Diagnosis of Chagas disease is hindered by discordance between screening and confirmatory test results for Trypanosoma cruzi infection. In periurban Arequipa, Peru, spatial analysis revealed that individuals with discordant test results are spatially clustered in hotspots of T. cruzi transmission, suggesting that discordant results likely represent true infections in this setting.

Chagas disease affects 8-11 million people throughout the Americas. Early detection is crucial for timely treatment and to prevent non-vectorial transmission. Recombinant antigen-based rapid tests had high sensitivity and specificity in laboratory evaluations, but no Peruvian specimens were included in previous studies. We evaluated Stat-Pak and Trypanosoma Detect rapid tests in specimens from Bolivia and Peru. Specimens positive by three conventional assays were confirmed positives; specimens negative by two or more assays were confirmed negatives. In Bolivian specimens, Stat-Pak and Trypanosoma Detect tests were 87.5% and 90.7% sensitive, respectively; both showed 100% specificity. Sensitivity in Peruvian specimens was much lower: 26.6-33.0% (Stat-Pak) and 54.3-55.2% (Trypanosoma Detect); both had specificities98%. Even in Bolivian specimens, these sensitivities are inadequate for stand-alone screening. The low sensitivity in Peru may be related to parasite strain differences. Chagas disease rapid tests should be field tested in each geographic site before widespread implementation for screening.

We standardized serodiagnosis of dogs infected with Trypanosoma cruzi using TESA (trypomastigote excreted-secreted antigen)-blot developed for human Chagas disease. TESA-blot showed 100% sensitivity and specificity. In contrast, ELISA using TESA (TESA-ELISA) or epimastigotes (epi-ELISA) as antigen yielded 100% sensitivity but specificity of 94.1% and 49.4%, respectively. When used in field studies in an endemic region for Chagas disease, visceral leishmaniasis and Trypanosoma evansi (Mato Grosso do Sul state, Central Brazil), positivities were 9.3% for TESA-blot, 10.7% for TESA-ELISA and 32% for epi-ELISA. Dogs from a non-endemic region for these infections (Rondonia state, western Amazonia) where T. cruzi is enzootic showed positivity of 4.5% for TESA-blot and epi-ELISA and 6.8% for TESA-ELISA. Sera from urban dogs from Santos, Sao Paulo, where these diseases are absent, yielded negative results. TESA-blot was the only method that distinguished dogs infected with T. cruzi from those infected with Leishmania chagasi and/or Trypanosoma evansi.

To examine the impact of tropical rain-forest destruction on malaria, we conducted a year-long study of the rates at which the primary malaria vector in the Amazon, Anopheles darlingi, fed on humans in areas with varying degrees of ecological alteration in the Peruvian Amazon. Mosquitoes were collected by human biting catches along the Iquitos-Nauta road at sites selected for type of vegetation and controlled for human presence. Deforested sites had an A. darlingi biting rate that was more than 278 times higher than the rate determined for areas that were predominantly forested. Our results indicate that A. darlingi displays significantly increased human-biting activity in areas that have undergone deforestation and development associated with road development.

Background: Millions of people are infected with Trypanosoma cruzi, the causative agent of Chagas disease in Latin America. Anti-trypanosomal drug therapy can cure infected individuals, but treatment efficacy is highest early in infection. Vector control campaigns disrupt transmission of T. cruzi, but without timely diagnosis, children infected prior to vector control often miss the window of opportunity for effective chemotherapy. Methods and Findings: We performed a serological survey in children 2–18 years old living in a peri-urban community of Arequipa, Peru, and linked the results to entomologic, spatial and census data gathered during a vector control campaign. 23 of 433 (5.3% [95% CI 3.4–7.9]) children were confirmed seropositive for T. cruzi infection by two methods. Spatial analysis revealed that households with infected children were very tightly clustered within looser clusters of households with parasite-infected vectors. Bayesian hierarchical mixed models, which controlled for clustering of infection, showed that a child’s risk of being seropositive increased by 20% per year of age and 4% per vector captured within the child’s house. Receiver operator characteristic (ROC) plots of best-fit models suggest that more than 83% of infected children could be identified while testing only 22% of eligible children. Conclusions: We found evidence of spatially-focal vector-borne T. cruzi transmission in peri-urban Arequipa. Ongoing vector control campaigns, in addition to preventing further parasite transmission, facilitate the collection of data essential to identifying children at high risk of T. cruzi infection. Targeted screening strategies could make integration of diagnosis and treatment of children into Chagas disease control programs feasible in lower-resource settings.