BACKGROUND Nyssorhynchus dunhami, a member of the Nuneztovari Complex, has been collected in Brazil, Colombia, and Peru and described as zoophilic. Although to date Ny. dunhami has not been documented to be naturally infected by Plasmodium, it is frequently misidentified as other Oswaldoi subgroup species that are local or regional malaria vectors. OBJECTIVES The current study seeks to verify the morphological identification of Nuneztovari Complex species collected in the peri-Iquitos region of Amazonian Peru, to determine their Plasmodium infection status, and to describe ecological characteristics of their larval habitats. METHODS We collected Ny. nuneztovari s.l. adults in 2011-2012, and Ny. nuneztovari s.l. larvae and adults in 2016-2017. When possible, samples were identified molecularly using cytochrome c oxidase subunit I (COI) barcode sequencing. Adult Ny. nuneztovari s.l. from 2011-2012 were tested for Plasmodium using real-time PCR. Environmental characteristics associated with Ny. nuneztovari s.l. larvae-positive water bodies were evaluated. FINDINGS We collected 590 Ny. nuneztovari s.l. adults and 116 larvae from eight villages in peri-Iquitos. Of these, 191 adults and 111 larvae were identified by COI sequencing; all were Ny. dunhami. Three Ny. dunhami were infected with P. falciparum, and one with P. vivax, all collected from one village on one night. Ny. dunhami larvae were collected from natural and artificial water bodies, and their presence was positively associated with other Anophelinae larvae and amphibians, and negatively associated with people living within 250m. MAIN CONCLUSIONS Of Nuneztovari Complex species, we identified only Ny. dunhami across multiple years in eight peri-Iquitos localities. This study is, to our knowledge, the first report of natural infection of molecularly identified Ny. dunhami with Plasmodium. We advocate the use of molecular identification methods in this region to monitor Ny. dunhami and other putative secondary malaria vectors to more precisely evaluate their importance in malaria transmission.

Objectives. To assess the competency of microscopists for malaria diagnosis using standardized slide sets in the Peruvian Amazon. Material and methods. Cross-sectional study carried out in 122 first level health facilities of the Peruvian Amazon, between July and September 2007. Within the frame of the project "Control Malaria in the border areas of the Andean Region: A community approach" (PAMAFRO), we evaluated the malaria diagnosis performance in 68 microscopists without expertise (< 1 year of expertise) and 76 microscopists with expertise (> 1 year) using standardized sets of 20 blood smear slides according to the World Health Organization (WHO) recommendations. A correct diagnosis (correct species identification) was defined as "agreement", a microscopist was qualified as an "expert" if they have an agreement ≥90% (≥ 18 slides with correct diagnosis), as a "referent" with an agreement between 80% and <90%, "competent" if they are between 70 and <80% and "in training" if they have <70%. Results. Microscopists with expertise (68.6%) had more agreement than those without expertise (48.2%). The competency assessment was acceptable (competent, referent, or experts levels) in 11.8% of the microscopists without expertise and in 52.6% from those with expertise. The agreement was lower using blood smear slides with P. falciparum with low parasitaemia, with P. malariae and with mixed infections. Conclusions. Is the first assessment, we found only one of three microscopists from the Peruvian Amazon is competent fro malaria diagnosis according to the WHO standards. From this baseline data, we have to continue working in order to improve the competency assessment of the microscopists within the frame of a quality assurance system

Objetivos. Evaluar la competencia de los microscopistas en el diagnóstico de la malaria mediante paneles de láminas estandarizados en la Amazonía peruana. Materiales y métodos. Estudio transversal, realizado entre los meses de julio y septiembre de 2007, en 122 establecimientos de salud de primer nivel de atención de la Amazonía peruana. En el marco del Proyecto PAMAFRO, se evaluó las competencias en el diagnóstico de malaria en 68 microscopistas sin experiencia (<un año experiencia) y 76 microscopistas con experiencia (>un año), utilizando paneles estandarizados de 20 láminas siguiendo las recomendaciones de la Organización Mundial de la Salud (OMS). Se definió concordancia como el porcentaje de aciertos que tuvo el microscopista en los diagnósticos (identificación de especie del parásito). Un microscopista con concordancia ≥90% (≥18 láminas acertadas) fue definido como "experto", 80 a <90% "referente", 70 a <80% "competente" y <70% "en entrenamiento". Resultados. Los microscopistas con experiencia (68,6%) tuvieron mayor concordancia que aquellos sin experiencia (48,2%). La competencia fue aceptable (niveles competente, referente o expertos) en 11,8% de los microscopistas sin experiencia y en 52,6% de aquellos con experiencia. La concordancia fue menor para láminas con P. falciparum con parasitemia baja, con P. malariae y con infecciones mixtas. Conclusiones. En esta primera evaluación, se evidenció que solo uno de cada tres microscopistas de la amazonía peruana es competente para el diagnóstico de malaria según estándares de la OMS. A partir de esta línea de base, se debe mejorar las competencias de los microscopistas en el marco de un sistema de aseguramiento de la calidad.

Objectives. To assess the competency of microscopists for malaria diagnosis using standardized slide sets in the Peruvian Amazon. Material and methods. Cross-sectional study carried out in 122 first level health facilities of the Peruvian Amazon, between July and September 2007. Within the frame of the project "Control Malaria in the border areas of the Andean Region: A community approach" (PAMAFRO), we evaluated the malaria diagnosis performance in 68 microscopists without expertise (< 1 year of expertise) and 76 microscopists with expertise (> 1 year) using standardized sets of 20 blood smear slides according to the World Health Organization (WHO) recommendations. A correct diagnosis (correct species identification) was defined as "agreement", a microscopist was qualified as an "expert" if they have an agreement ≥90% (≥ 18 slides with correct diagnosis), as a "referent" with an agreement between 80% and <90%, "competent" if they are between 70 and <80% and "in training" if they have <70%. Results. Microscopists with expertise (68.6%) had more agreement than those without expertise (48.2%). The competency assessment was acceptable (competent, referent, or experts levels) in 11.8% of the microscopists without expertise and in 52.6% from those with expertise. The agreement was lower using blood smear slides with P. falciparum with low parasitaemia, with P. malariae and with mixed infections. Conclusions. Is the first assessment, we found only one of three microscopists from the Peruvian Amazon is competent fro malaria diagnosis according to the WHO standards. From this baseline data, we have to continue working in order to improve the competency assessment of the microscopists within the frame of a quality assurance system

Objetivos. Evaluar la competencia de los microscopistas en el diagnóstico de la malaria mediante paneles de láminas estandarizados en la Amazonía peruana. Materiales y métodos. Estudio transversal, realizado entre los meses de julio y septiembre de 2007, en 122 establecimientos de salud de primer nivel de atención de la Amazonía peruana. En el marco del Proyecto PAMAFRO, se evaluó las competencias en el diagnóstico de malaria en 68 microscopistas sin experiencia (<un año experiencia) y 76 microscopistas con experiencia (>un año), utilizando paneles estandarizados de 20 láminas siguiendo las recomendaciones de la Organización Mundial de la Salud (OMS). Se definió concordancia como el porcentaje de aciertos que tuvo el microscopista en los diagnósticos (identificación de especie del parásito). Un microscopista con concordancia ≥90% (≥18 láminas acertadas) fue definido como "experto", 80 a <90% "referente", 70 a <80% "competente" y <70% "en entrenamiento". Resultados. Los microscopistas con experiencia (68,6%) tuvieron mayor concordancia que aquellos sin experiencia (48,2%). La competencia fue aceptable (niveles competente, referente o expertos) en 11,8% de los microscopistas sin experiencia y en 52,6% de aquellos con experiencia. La concordancia fue menor para láminas con P. falciparum con parasitemia baja, con P. malariae y con infecciones mixtas. Conclusiones. En esta primera evaluación, se evidenció que solo uno de cada tres microscopistas de la amazonía peruana es competente para el diagnóstico de malaria según estándares de la OMS. A partir de esta línea de base, se debe mejorar las competencias de los microscopistas en el marco de un sistema de aseguramiento de la calidad.

Leishmania (Viannia) peruviana was isolated from 1/75 Lutzomyia peruensis captured during May 2006 in an endemic cutaneous leishmaniasis region of the Peruvian Andes (Chaute, Huarochiri, Lima, Peru). Sand fly gut with promastigotes was inoculated into a hamster and the remaining body was fixed in ethanol. L. (Viannia) sp. was determined by polymerase chain reaction (PCR), and Leishmania species through molecular genotyping by PCR-restriction fragment length polymorphism analyses targeting the genes cpb and hsp70, resulting L. (V.) peruviana. The infected sand fly appeared 15 days after the rains finished, time expected and useful real time data for interventions when transmission is occurring.