Black Fly (Diptera: Simuliidae) Assemblages of High Andean Rivers Respond to Environmental and Pollution Gradients.

Simuliid larvae are common inhabitants of mountain rivers throughout the world, where they can serve as ecological indicators. Black flies were sampled during three seasons in four rivers in the upper basin of the Bogotá River in the Colombian Andes, and physical, chemical, and hydrological data were recorded. Multivariate methods were used to determine the relationships between the presence and abundance of simuliid species and environmental characteristics. Fourteen species were found: eight in the genus Gigantodax (Enderlein, Diptera, Simuliidae) and six in the genus Simulium (Latreille, Diptera, Simuliidae). Dissolved oxygen, dissolved solids, redox potential, chemical oxygen demand, and nutrients contributed to an explanation of species distributions. Species in clean waters and in more polluted waters had narrow niches; those in low to moderately polluted waters had broader niches. Species in the lower reaches of the watercourses had greater turnover, perhaps because the most sensitive species had disappeared and been replaced by more tolerant species. Thus, simuliids can be used as predictors of environmental characteristics of Andean rivers and can be useful in the evaluation and management of these watersheds.

Cellular and Humoral Responses to Cte f 2, a Cat Flea Allergen, in Children with Papular Urticaria.

BACKGROUND: Papular urticaria (PU) is a common insect bite skin hypersensitivity in tropical countries. In order to gain insight into its causal allergens, we aimed to evaluate cellular and humoral immune responses to the recombinant salivary antigen Cte f 2 from the cat flea Ctenocephalides felis. METHOD: Sixty patients with PU and 27 healthy controls were included in this study. Specific IgE, IgG, IgG1, and IgG4 against Cte f 2 and C. felis extract were determined by ELISA. The T-cell response was analyzed using a carboxyfluorescein succinimidyl ester (CFSE)-based dilution assay and Th1/Th2/Th17 cytokine measurements. In addition, a proteomic analysis of IgG and IgE reactive spots of C. felis extract was performed. RESULTS: The frequency of IgE sensitization to Cte f 2 was similar between patients (36.7%) and controls (40.7%). The specific IgE, IgG1, and IgG4 responses to Cte f 2 and C. felis extract were not significantly different between patients and controls. Among the 3 conditions (i.e., Cte f 2, C. felis extract, and only medium) Cte f 2 was the strongest inducer of CD3+CD4+ proliferation in the patients; however, the mean response was not significantly different from those in controls (Cte f 2: 4.5 vs. 2.5%; p = 0.46). No salivary proteins were identified in C. felis, and most of the spots were identified as muscle-skeletal components (tropomyosin, actin, myosin, and ankirin). CONCLUSIONS: Cte f 2 induces IgE and IgG production as well as T-cell proliferation in children living in a geographical area where PU induced by a flea bite is common. The use of C. felis extract is not recommended for the study of bite-induced hypersensitivity disease since salivary antigens are not well represented.

Distribution of black flies (Diptera: Simuliidae) along an elevational gradient in the Andes Mountains of Colombia during the El Niño Southern Oscillation.

Vector ecology is a key factor in understanding the transmission of disease agents, with each species having an optimal range of environmental requirements. Scarce data, however, are available for how interactions of local and broad-scale climate phenomena, such as seasonality and the El Niño Southern Oscillation (ENSO), affect simuliids. We, therefore, conducted an exploratory study to examine distribution patterns of species of Simuliidae along an elevational gradient of the Otún River in the Colombian Andes, encompassing four ecoregions. Larval and pupal simuliids were sampled at 52 sites ranging from 1800 to 4750 m above sea level in dry and wet seasons and during the La Niña phase (2011-2012) and the El Niño phase (2015-2016) of the ENSO; physicochemical measurements were taken during the El Niño phase. Twenty-seven species in two genera (Gigantodax and Simulium) were collected. Species richness and occurrence in each ecoregion were influenced by elevation, seasonality, and primarily the warm El Niño and cool La Niña phases of the ENSO. The degree of change differed among ecoregions and was related to physicochemical factors, mainly with stream discharge. Some putative simuliid vectors of Leucocytozoon, such as G. misitu and S. muiscorum, markedly changed in distribution and occurrence, potentially influencing parasite transmission.

Potential pharmacological use of salivary compounds from hematophagous organisms / Uso potencial farmacológico de los compuestos salivares de organismos hematófagos

Abstract Introduction: The saliva of hematophagous or blood-sucking insects contains different substances that allow obtaining and ingesting the blood of their vertebrate hosts without being detected. Objective: To explore the salivary compounds of hematophagous insects which have vasodilator, anticoagulant, anti-inflammatory, immunomodulatory and anesthetic properties, and that can be exploited due to their high pharmacological potential. Materials and methods: A non-systematic literature review was done in PubMed, EMBASE, and ScienceDirect OvidSP; data was not limited by date, language nor item type. Articles on salivary compounds of blood-sucking insects, whose main topic was the effects on hemostasis, immunomodulation and drug use, were sought. 59 articles met the criteria for inclusion in the review. Conclusions: The saliva of hematophagous insects has a wide variety of molecules that constitute a source of research and have an incalculable potential for the discovery of compounds that could be pharmacologically useful.

Resumen Introducción. La saliva de los artrópodos hematófagos contiene un arsenal de compuestos que les permite acceder a la sangre de sus hospederos vertebrados sin ser detectados. Objetivo. Explorar los compuestos salivares de insectos hematófagos que tienen propiedades vasodilatadoras, anticoagulantes, antiinflamatorias, inmunomoduladoras y anestésicas, las cuales se pueden aprovechar por su alto potencial farmacológico. Materiales y métodos. Se realizó una revisión no sistemática de la literatura mediante búsqueda electrónica en las bases de datos PubMed, EMBASE, OvidSP y ScienceDirect; la búsqueda no se limitó por fecha, idioma ni tipo de artículo. Se buscaron artículos sobre los compuestos salivares de los insectos hematófagos, cuyo tema central fuese los efectos en la hemostasia, inmunomodulación y uso farmacológico. Se encontraron 59 artículos que cumplían con los criterios para ser incluidos en la revisión. Conclusión. La saliva de los insectos hematófagos posee gran variedad de moléculas, lo que ofrece una fuente de investigación y un potencial incalculable para el descubrimiento de compuestos que podrían llegar a tener utilidad farmacológica.

Combination of five diagnostic tests to estimate the prevalence of hookworm infection among school-aged children from a rural area of colombia.

BACKGROUND: Public health programs for the control of soil-transmitted helminthiases require valid diagnostic tests for surveillance and parasitic control evaluation. However, there is currently no agreement about what test should be used as a gold standard for the diagnosis of hookworm infection. Still, in presence of concurrent data for multiple tests it is possible to use statistical models to estimate measures of test performance and prevalence. The aim of this study was to estimate the diagnostic accuracy of five parallel tests (direct microscopic examination, Kato-Katz, Harada-Mori, modified Ritchie-Frick, and culture in agar plate) to detect hookworm infections in a sample of school-aged children from a rural area in Colombia. METHODS AND RESULTS: We used both, a frequentist approach, and Bayesian latent class models to estimate the sensitivity and specificity of five tests for hookworm detection, and to estimate the prevalence of hookworm infection in absence of a Gold Standard. The Kato-Katz and agar plate methods had an overall agreement of 95% and kappa coefficient of 0.76. Different models estimated a sensitivity between 76% and 92% for the agar plate technique, and 52% to 87% for the Kato-Katz technique. The other tests had lower sensitivity. All tests had specificity between 95% and 98%. The prevalence estimated by the Kato-Katz and Agar plate methods for different subpopulations varied between 10% and 14%, and was consistent with the prevalence estimated from the combination of all tests. The Harada-Mori, Ritchie-Frick and direct examination techniques resulted in lower and disparate prevalence estimates. Bayesian approaches assuming imperfect specificity resulted in lower prevalence estimates than the frequentist approach.

Leucocytozoon Diversity and Possible Vectors in the Neotropical highlands of Colombia.

Studies of the lowland avifauna in the Neotropical Region have shown a paucity of Leucocytozoon species. However, surveys conducted in the Colombian highlands revealed a great diversity of these parasites infecting resident birds. To further investigate the relationship between Leucocytozoon diversity, the potential vectors, and altitudinal distribution, birds from 41 families were sampled at low and high elevations in Colombia. Blood samples were screened by microscopy, and a fragment of cytochrome b was amplified from Leucocytozoon-positive samples. The complete mitochondrial genome was also obtained for each morphospecies of Leucocytozoon. Leucocytozoon species were detected in resident birds, with various degrees of host specificity, at elevations from 2,400 to 3,950 meters above sea level, where five new host-parasite associations were discovered. Phylogenetic analysis based on the cytochrome b fragment suggested that two nominal taxa, L. fringillinarum and L. majoris, are species complexes. Blood sources of Simuliidae revealed generalist-feeding habits that included avian and mammalian hosts. Molecular analysis of parasites in black flies indicated a close relationship with the parasites found in birds. Our investigation provides further evidence that the distribution and transmission of Leucocytozoon species in the Neotropics are influenced by elevation, with the highest prevalence between 2,400 and 3,200 m asl.

Haemoproteus erythrogravidus n. sp. (Haemosporida, Haemoproteidae): Description and molecular characterization of a widespread blood parasite of birds in South America.

The great diversity of birds and ecosystems in the Andean mountains has been understudied in terms of their parasite species. We describe a new Haemoproteus parasite, H. (Parahaemoproteus) erythrogravidus infecting Zonotrichia capensis (Rufous-Collared Sparrow) in South America. The description of this blood parasite species is supported by morphological and molecular data based on a fragment of cytochrome b gene (cyt b) and complete mitochondrial genome sequences. The new species is closely related to H. (Parahaemoproteus) coatneyi, and it can be readily distinguished from the latter parasite due to morphology of its blood stages, particularly 1) the formation of a marked protrusion on envelope of infected erythrocytes by the majority of developing gametocytes, a feature which is unique for this Haemoproteus species and 2) the extremely attenuated width of the growing dumbbell-shaped macro- and microgametocytes. Additionally, Haemoproteus erythrogravidus is shown to be a monophyletic taxon that diverges from Haemoproteus coatneyi at the molecular level. We provide the complete mitochondrial DNA genome for both H. coatneyi and H. erythrogravidus. Molecular and morphological evidences indicate that H. erythrogravidus is present in Ecuador and Colombia, and genetic lineages with 100% of identity for the cyt b gene were reported in Chile, Perú, and Venezuela. Our study also indicates that H. erythrogravidus and H. coatneyi are sympatric sister taxa sharing Z. capensis as a host species across its distribution, which could be the result of sympatric speciation or complex biogeographic processes. Further studies on the distribution and evolutionary history of Z. capensis and its parasites H. erythrogravidus and H. coatneyi insight for our better understanding of the factors and dynamics driving parasite speciation.

Molecular Epidemiology of Entamoeba: First Description of Entamoeba moshkovskii in a Rural Area from Central Colombia.

BACKGROUND: Entamoeba histolytica, E. dispar and E. moshkovskii are the most frequent species described in human infection where E. histolytica is the only true pathogen. The epidemiology of this infection is complex due to the absence of a routine exam that allows a correct discrimination of the Entamoeba species complex. Therefore, molecular methods appear as the unique epidemiological tool to accomplish the species discrimination. Herein, we conducted a cross-sectional study to determine the frequency of Entamoeba species infections in a group of asymptomatic individuals from a rural area in central Colombia. METHODOLOGY/PRINCIPAL FINDINGS: A total of 181 fecal samples from asymptomatic children under 16 years old from the hamlet La Vírgen, Cundinamarca (Colombia) that voluntarily accepted to participate in the study were collected. The fecal samples were examined by light microscopy and DNA-extracted, subsequently submitted to molecular discrimination of E. dispar/E. histolytica/E. moshkovskii infection based on a multiplex PCR assay targeting the 18S rRNA fragment. To confirm the species description, twenty samples were randomly submitted to DNA sequencing of the aforementioned fragment. By direct microscopic examination, frequency of the complex E. histolytica/E. dispar/E. moshkovskii was 18.8% (34/181). PCR showed a frequency of 49.1% (89/181), discriminated as 23.2% (42/181) that were positive for E. dispar, 25.4% (46/181) for E. moshkovskii and 0.55% (1/ 181) for E. histolytica. Also, mixed infections were detected between E. dispar and E. moshkovskii at 4.42% (8/181) of the samples. Molecular barcoding confirmed the diagnosis depicted by the multiplex PCR assay. CONCLUSIONS/SIGNIFICANCE: This is the first description of E. moshkovskii in Colombia and the second report in South-America to our knowledge. Our results suggest the need to unravel the true epidemiology of Entamoeba infections around the world, including the real pathogenic role that E. moshkovskii may have.

Implementación y evaluación de una estrategia combinada de educación en salud y quimioterapia masiva para el control de las geohelmintiasis, en una zona rural de Colombia / Implementation and evaluation of a combined strategy using health education and massive chemotherapy to control geohelminthiasis in a rural zone in Colombia

Introducción: Las geohelmintiasis son infecciones prevalentes en los países en vía de desarrollo, que afectan principalmente a la población escolar. Las estrategias de control que combinan quimioterapia masiva con educación en salud, pueden producir reducciones significativas de estas infecciones, sostenibles en el tiempo. En este estudio se implementó y evaluó, una intervención integral basada en una estrategia lúdica de educación en salud, más quimioterapia masiva en una zona rural de Colombia. Metodología: Diseño cuasi-experimental, realizado en La Virgen, Cundinamarca (Colombia). Toda la población escolar fue sometida a la estrategia educativa en salud, acompañada de tratamiento semestral con albendazol, entre finales del 2010 y mediados del 2013. La muestra analítica fue constituida por 119 niños (mediana= 11; Rango Intercuartílico = 8-13 años). Se evaluaron los cambios anuales en la presencia de geohelmintos usando pruebas no paramétricas para muestras relacionadas y modelos de regresión logística de efectos fijos. Resultados: En la medición basal, las prevalencias de los geohelmintos fueron: 22,28% (IC 95% 16,36%-28,20%) para Ascaris lumbricoides; 37,31% (IC 95% 30,42%-44,19%) para Trichuris trichiura y 13,47% (IC 95% 8,61%-18,33%) para Uncinaria sp. Para la medición final, estas prevalencias cayeron significativamente a 5,30% (IC 95% 1,68%-8,91%) para A. lumbricoides; 16,55% (IC 95% 10,56%-22,55%) para T. trichiura y 4,64% (IC95% 1,42%-8,03%) para Uncinaria sp. Conclusión: El presente estudio, pese a que no constituye una evaluación real de impacto, provee una experiencia aparentemente exitosa de implementación de una estrategia integral para el control de las geohelmintiasis, que podría servir como modelo para otras intervenciones en áreas rurales del país.

Introduction: Geohelminthiasis is an infection which is prevalent in developing countries, and primarily affects schoolchildren. Control strategies that combine massive chemotherapy with health education can produce significant reductions in this infection that are sustainable over time. This study implemented and evaluated a comprehensive intervention combining a health education strategy and massive chemotherapy in a rural zone in Colombia. Methodology: Quasi-experimental design performed in La Virgen, Cundinamarca (Colombia). The entire school population participated in the health education strategy, in conjunction with biannual treatments with albendazole, between late 2010 and the middle of 2013. The analytical sample consisted of 119 children (median = 11, interquartile range = 8-13 years of age). Yearly changes in the presence of geohelminthes were evaluated using non-parametric tests for related samples and fixed-effect logistic regression models. Results: Prevalences of geohelminthes at baseline were: 22,28% (CI 95% 16,36%-28,20%) for Ascaris lumbricoides; 37,31%( CI 95% 30,42%-44,19%) for Trichuris trichiura and 13,47% (CI 95% 8,61%-18,33%) for Uncinaria sp. The final measurements of these prevalences decreased to 5,30%(CI 95% 1,68%-8,91%) for A. lumbricoides; 16,55% (CI 95% 10,56%-22,55%) for T. trichiura and 4,64% (CI 95% 1,42%-8,03%) for Uncinaria sp. Conclusions: Although the present study does not constitute an actual impact evaluation, it provides an apparently successful experience with implementing a comprehensive strategy to control geohelminthiasis, which could serve as a model for other interventions in rural areas in the country.

Molecular diagnosis and genotype analysis of Giardia duodenalis in asymptomatic children from a rural area in central Colombia.

Giardiasis is a parasitic infection that affects around 200 million people worldwide. This parasite presents a remarkable genetic variability observed in 8 genetic clusters named as 'assemblages' (A-H). These assemblages are host restricted and could be zoonotic where A and B infect humans and animals around the globe. The knowledge of the molecular epidemiology of human giardiasis in South-America is scarce and also the usefulness of PCR to detect this pathogen in fecal samples remains controversial. The aim of this study was to conduct a cross-sectional study to compare the molecular targets employed for the molecular diagnosis of Giardia DNA and to discriminate the parasite assemblages circulating in the studied population. We analyzed 181 fecal samples from Children at La Virgen, Cundinamarca, Colombia that were DNA-extracted and analyzed by SSU rDNA, tpi and gdh loci. We observed positivity by microscopy of 13% and by PCR around 76-80% depending on the molecular marker. Additionally, a lack of statistical concordance between microscopy and PCR was detected. Regarding the genetic assemblages, we detected assemblage A (3%), assemblage B (90%) and mixed infections assemblages A+B (7%). Hence, the sub-assemblages were typed as AI, AII, BIII and BIV across the population. This study represents a reliable attempt to understand the molecular epidemiology of giardiasis in Colombia and the use of PCR to detect cryptic infections. The epidemiological implications are herein discussed.

Agreement of the Kato-Katz test established by the WHO with samples fixed with sodium acetate analyzed at 6 months to diagnose intestinal geohelminthes.

The aim of this study was to evaluate the performance of the Kato-Katz test (WHO version) with stool samples from a rural area, fixed with sodium acetate (SAF). The Kato-Katz test was used to compare unfixed samples (conventional test) with the same samples containing SAF fixative at time 0 and at 6 months. The study included stools from 154 subjects. A marginally statistically significant decrease in prevalence was estimated only for hookworm, when comparing unfixed samples versus the SAF fixed samples read at 6 months (p=0.06). A significant reduction in parasite load was found for hookworm (p<0.01) and Trichuris trichiura (p<0.01) between the unfixed and the fixed sample read at 6 months, but not for Ascaris lumbricoides (p=0.10). This research suggests that the SAF fixative solution is a good option for transporting samples for diagnosis, especially in rural areas in developing countries.

Avian haemosporidians from Neotropical highlands: Evidence from morphological and molecular data.

Avian haemosporidian parasites have been scarcely studied in the Neotropical highlands despite the high avian diversity reported and the uniqueness of these ecosystems. The aims of this study were to examine Haemoproteus and Plasmodium diversity based on morphological and molecular data, as well as to explore the concordance between these two approaches, when identifying species. We sampled 1487 birds belonging to 166 species, in localities of the Colombian Andean region at elevations ranging from 2100 to 4000 m above sea level. Here, we report twelve morphological parasite species, of which five are undescribed. Thirty parasite cytochrome b lineages are reported, 17 of which for the first time. We provide morphological information and illustrations, as well as, cytochrome b lineages for six morphospecies: Haemoproteus columbae, Haemoproteus witti, Haemoproteus coatneyi, Haemoproteus vireonis, Plasmodium lutzi, and Plasmodium unalis. This is the first report to provide a linkage between morphology and a molecular lineage for H. witti. Cytochrome b gene proved to be useful for species determination as DNA barcoding. Differences in parasite composition between lowlands and highlands in Colombia suggest a replacement of avian Plasmodium fauna. Parasite lineages restricted to either Colombian resident or Nearctic migratory birds were found; but a single lineage common in both has not been recorded in Nearctic non-migratory birds. We generated valuable information by using both morphological and molecular data representing competent host-parasite relationships which are based on observation of gametocytes in circulation; and increased the taxon sampling of avian haemosporidian.

Integrated taxonomy of a new species of black fly in the subgenus Trichodagmia (Diptera: Simuliidae) from the Páramo Region of Colombia.

A new species of simuliid from the Andean Mountains of Colombia is described on the basis of females, males, pupae, larvae, polytene chromosomes, and COI and ITS2 DNA sequences. Simulium (Trichodagmia) chimguazaense new species is structurally, chromosomally, and molecularly distinct from its nearest relatives, S. muiscorum Bueno, Moncada & Muñoz de Hoyos and S. sumapazense Coscarón & Py-Daniel. 

Comparison of hemocytes of V-instar nymphs of Rhodniusprolixus (Stål) and Rhodnius robustus (Larousse 1927), before and after molting / Comparación de los hemocitos de las ninfas de V instar de Rhodnius prolixus (Stål) and Rhodnius robustus, antes y después de la muda (Larousse 1927)

Background. The immune response of insects involves humoral factors and cellular elements known as hemocytes. There are different reports that explore the response of hemocytes to infections, but the effect that molting has on this response has not been explored so far. We hypothesized that there would be a change in the percentage of hemocytes as a response to the molting process. Objective. The aim of this work was to compare the hemogram (CBC), the formula, and the differential count of hemocytes in IV instar before molting and in V instar nymphs 24 hours after molting in two species: Rhodnius prolixus and Rhodnius robustus. Materials and methods. We assayed different staining methodologies including Giemsa, Alcian Blue pH 2.0, Alcian Blue pH 2.6, Gomori substrate, PAS (Schiff), Sudan Black and Papanicolau with positive controls for each one. In the Gomori staining, we observed lysosomes in the granulocytes and plasmatocytes, but the differentiation was better detected using Giemsa staining. Results. There were no statistically significant differences between the two species studied in plasmatocytes (p=0,053) or even in granulocytes (p=0,5). However, differences were significant in the prohemocytes (p=0,001) during the molting process in both Rhodnius prolixus and Rhodnius robustus. Conclusions. Significant differences in prohemocytes between nymphs of IV and V instar were detected. No significant differences in the amount of cells were observed between the two species and the two stages. These findings may be explained due to their role as precursor cell of prohemocytes.

Antecedentes. La respuesta inmune de los insectos involucra factores humorales y elementos celulares llamados hemocitos. Existen varios trabajos explorando la respuesta de los hemocitos frente a infecciones, pero no como respuesta al proceso de muda. Objetivo. Comparar el hemograma: la fórmula y el recuento diferencial de hemocitos de Rhodnius prolixus y Rhodnius robustus en ninfas de IV estadio antes de la muda y ninfas de V estadio 24 horas después de la muda. Materiales y métodos. Se ensayaron las coloraciones Giemsa, Alcian Blue pH 2.0, Alcian Blue pH 2.6, sustrato Gomori, PAS (Schiff), Sudán Negro, Papanicolau, con controles positivos para cada una. Con la coloración de Gomori se observaron lisosomas en los granulocitos y en plasmatocitos, pero la diferenciación se logró mejor con la coloración de Giemsa. Resultados. Los plasmatocitos fueron más abundantes en las ninfas de IV estadio de Rhodnius robustus y en las de V estadio de las dos especies, pero en las ninfas de V estadio de R. prolixus, el recuento de prohemocitos fue mayor; para los oenocitoides, esferulocitos y adipohemocitos el recuento no alcanzó al 1 %. En las dos especies no se encontraron diferencias estadísticamente significativas en plasmatocitos (p=0,05319), ni en granulocitos (p=0,5), entre las ninfas de IV y V estadio, pero fue significativa en prohemocitos (p=0,001). Conclusiones. Se detectaron diferencias significativas en prohemocitos entre las ninfas de IV y V estadio. No hubo un aumento significativo en el conteo de células en los diferentes estadios.

COMPORTAMIENTO DE OVIPOSICIÓN Y CARACTERIZACIÓN DE LA MICROESTRUCTURA CORIONICA DE LOS HUEVOS DE Simulium dinelliii (Joan) (Diptera: Simuliidae) / Oviposition Behavior and Microstructure of the Eggs of Simulium dinellii (Joan) (Diptera: Simuliidae)

Little is known about the oviposition habits and egg structure of Neotropical members of the Simuliidae family. The oviposition behavior of Simulium dinellii (Joan) was observed near at Otún Quimbaya, Colombia. Small groups of females hovered about 2 cm above the water surface to oviposit. Some of these females had been capture and the eggs were obtained. These eggs were counted, measured and processed by scanning electron microscopy. Subtriangular eggs had a latticework of polygons on the endochorionic surface. Chorionic differences between the eggs of S. dinellii and the few previously studied species suggest potential taxonomic value of egg microstructure.

El conocimiento sobre los hábitos de oviposición y estructura de los huevos de la familia Simuliidae es muy escaso. En este estudio se observó el comportamiento de oviposición de Simulium dinellii (Joan) en quebradas cercanas al Santuario de fauna y flora Otún Quimbaya, Colombia. El cual consistió en la formación de pequeños grupos de hembras que sobrevolaban a 2 cm de la superficie del agua para ovipositar, algunas de estas hembras fueron colectadas, obteniendo así los huevos, los cuales fueron contados, medidos y procesados por Microscopia Electrónica de Barrido con el fin de caracterizar la estructura endocoriónica. Los huevos presentaron una forma subtriangular, con una entramado poligonal sobre la superficie. Las características morfológicas observadas en los huevos de S. dinellii fueron diferentes a las previamente reportadas en otras especies de la familia Simuliidae, sugiriendo un posible valor taxonómico de la microestructura del huevo.

Leucocytozoon pterotenuis sp. nov. (Haemosporida, Leucocytozoidae): description of the morphologically unique species from the Grallariidae birds, with remarks on the distribution of Leucocytozoon parasites in the Neotropics.

We describe morphologically unique Leucocytozoon pterotenuis sp. nov. (Haemosporida, Leucocytozoidae), the first reported leucocytozoid species developing in fusiform host cell found in a Neotropical passeriform bird. The type host of this parasite is the Chestnut-crowned Antpitta (Grallaria ruficapilla, Grallariidae), an elusive native passerine bird whose natural history remains, to a large degree, unexplored. This bird was captured in Palacio forest in the damping zone of Chingaza National Natural Park, Cundinamarca, Colombia, at 2900 m above sea level (asl). Gametocytes of the new species develop both in roundish and fusiform host cells. This parasite is readily morphologically distinguishable from the described Leucocytozoon species because its host cells possess the narrow (needle-like) spindle-shaped processes, which length markedly exceeds their width. Additionally, the host cell nucleus markedly extends into the processes. Phylogenetic relationships were constructed based on a fragment of the mitochondrial cytochrome b gene and the complete mitochondrial genome. Phylogenetic analysis placed the lineage of L. pterotenuis in different positions depending on the length of the sequence analyzed that is likely due to poor sampling of Leucocytozoon species, especially from rare or non-passerine hosts, as well as a paucity of complete mitochondrial sequences of these parasites. Available data indicate that Leucocytozoon parasites are distributed mainly in mountain regions of the Neotropics where unique morphological forms have been recently discovered. To a better knowledge of the diversity of Leucocytozoon spp. and their host-vector-parasite interactions in Neotropical countries, additional deep and intensive samplings are needed, particularly in orders different to Passeriformes.

Detection of Strongyloides stercoralis in Tierralta, Colombia using four parasitological methods / Detección de Strongyloides stercoralis en Tierralta, Colombia, utilizando cuatro métodos parasitológicos

Introduction: soil-borne helminth Strongyloides stercoralis is one of the most neglected among neglected tropical diseases. A study was conducted of the presence of S. stercoralis in a village from the department of Córdoba, Colombia, with the purpose of comparing the effectiveness of several diagnostic methods. Methods: stool samples from 262 persons were evaluated. Each sample was examined with four parasitological techniques: direct examination, agar plate culture (APC), the modified Baermann method, and the Harada-Mori technique. Results: S. stercoralis was detected by at least one of the techniques in four of the 262 samples: the Harada-Mori technique detected 2 cases, APC 1 case and direct examination 1 case. The modified Baermann method did not detect any case. No significant differences were found when comparing the techniques. Conclusions: results show that S. stercoralis is not endemic in the village of Córdoba, and that parasitological techniques should be used in combination to improve the quality of diagnosis.

Introducción: el helminto transmitido por el suelo, Strongyloides stercoralis es uno de los más olvidados entre las enfermedades tropicales desatendidas. Estudiamos la presencia de S. stercoralis en un pueblo en el departamento de Córdoba, Colombia, y evaluamos comparativamente el desempeño de diferentes métodos diagnósticos. Métodos: se evaluaron muestras de heces tomadas de 262 personas; cada muestra fue examinada usando cuatro técnicas parasitológicas: examen directo, método de agar en placa (APC), la técnica de Baermann modificado y el método de Harada-Mori. Resultados: S. stercoralis se detectó en cuatro de las 262 muestras evaluadas por al menos una de las técnicas utilizadas; el método deHarada-Mori detectó 2 casos, APC 1 caso y el examen directo 1 caso, mientras que la técnica de la Baermann modificado no detectó casos. No hubo diferencias significativas al comparar las técnicas. Conclusiones: estos resultados permiten concluir que S. stercoralis no es endémico en el pueblo de Córdoba y que las técnicas parasitológicas deben ser combinadas para mejorar el diagnóstico.

Description of Leucocytozoon quynzae sp. nov. (Haemosporida, Leucocytozoidae) from hummingbirds, with remarks on distribution and possible vectors of leucocytozoids in South America.

We describe Leucocytozoon quynzae sp. nov. (Haemosporida, Leucocytozoidae), which is the first Leucocytozoon parasite identified to species level in hummingbirds. It was found in the Amethyst-throated Sunangel (Heliangelus amethysticollis, Trochilidae, Apodiformes) captured in the Palacio Forest, which belongs to the damping zone of Chingaza National Natural Park, Cundinamarca, Colombia, at 2,900 m above sea level where the transmission occurs; the new species were found both in the high Andean forest and Paramo ecosystem. This parasite is described based on the morphology of its blood stages, a fragment of the mitochondrial cytochrome b gene, and the complete mitochondrial genome. Illustrations of blood stages of the new species are given, and the phylogenetic analysis places this lineage in a well-supported clade with other lineages of unidentified to species level leucocytozoids reported in the Trochilidae birds elsewhere. The new species possess gametocytes in roundish host cells; it can be readily distinguished from other similar leucocytozoids, primarily due to (1) a comma-like shape of the host cell nucleus, which extended one half or less of the circumference of the gametocyte and (2) a large number of prominent volutin granules in the cytoplasm. Identical mitochondrial cytochrome b sequence of Leucocytozoon quynzae was found in different hummingbird species at the type locality and also was reported in one passerine bird at the highlands of Peru. Leucocytozoon quynzae is the first leucocytozoid parasite described from South American birds; its transmission occurs both at low temperatures and high elevations. We discuss some patterns of distribution of avian leucocytozoids in South America and the role of Gigantodax spp. (Diptera, Simuliidae) as potential vectors of Leucocytozoon parasites in the Andean Region.

Description and molecular characterization of Plasmodium (Novyella) unalis sp. nov. from the Great Thrush (Turdus fuscater) in highland of Colombia.

Plasmodium (Novyella) unalis sp. nov. was found in the Great Thrush, Turdus fuscater (Passeriformes, Turdidae) in Bogotá, Colombia, at 2,560 m above sea level where the active transmission occurs. This parasite is described based on the morphology of its blood stages and a fragment of the mitochondrial cytochrome b gene (lineage UN227). Illustrations of blood stages of new species are given, and the phylogenetic analysis identifies closely related species and lineages of avian malaria parasites. The new species is most similar to Plasmodium (Novyella) vaughani (lineage SYAT05), a cosmopolitan avian malaria parasite; these parasites are also closely related genetically, with a genetic difference of 3.2% between them. P. unalis can be readily distinguished from the latter species morphologically, primarily due to the (1) presence of a single large, circular shaped pigment granule in the erythrocytic trophozoites and meronts; (2) presence of prominent vacuoles in trophozoites and growing meronts; and (3) presence of predominantly fan-like shaped erythrocytic meronts. Cytochrome b lineages with high similarity to the new species have been reported in Costa Rica, Brazil, Chile, and USA. It is probable that the new species of malaria parasite is widely distributed in the New World. This parasite has been reported only in the Great Thrush at the study site and might have a narrow range of avian hosts. Records of P. unalis are of particular theoretical interest due to its active transmission at highlands in Andes. Possible influence of urbanization on transmission of this malaria parasite in Bogotá is discussed.