Phlebotomine (Diptera: Psychodidae) species and their blood meal sources in a new leishmaniasis focus in Los Montes de María, Bolívar, in northern Colombia / Especies de flebotomíneos y sus fuentes de ingesta sanguínea en un nuevo foco de leishmaniasis en Los Montes de María (Bolívar) al norte de Colombia.

Introduction. El Alférez, a village in Los Montes de María (Bolívar, Colombia) and a macro-focus of leishmaniasis, recorded its first case in 2018, evidencing changes in the distribution and eco-epidemiology of the disease, although interactions between vectors and local fauna remain unknown. Objective. To evaluate the diversity of sandflies and their blood meal sources in the community of El Alférez in the municipality of El Carmen de Bolívar (Bolívar, Colombia). Materials and methods. In 2018, sandflies were collected using LED-based light traps in domestic, peridomestic, and sylvatic ecotopes and identified at the species level. Multiplex polymerase chain reaction targeting the mitochondrial cytochrome B gene was used to analyze blood from the digestive tract. Results. Lutzomyia evansi was the most abundant species (71.85%; n = 485/675), followed by Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci, and Lu.aclydifera. Twenty-five percent of the species had blood meals from Canis familiaris (36.00%; n = 9/25), Ovis aries (36.00%; n=9:/25), Bos taurus (24.00%; n = 6/25), Sus scrofa (20.00%; n = 5/25), and Homo sapiens (8.00%; n = 2/25). Lutzomyia evansi registered the highest feeding frequency (68.00%; n = 17/25), predominantly on a single (44.00%; n = 11/25) or multiple species (24.00%; n = 6/25). Conclusion. Results indicate a eclectic feeding behavior in Lu. evansi, implying potential reservoir hosts for Leishmania spp. and increasing transmission risk. This study is a first step towards understanding the diversity of mammalian blood sources used by sandflies, that may be crucial for vector identification and formulation of effective control measures.

Introducción. En 2018, en la vereda El Alférez de Los Montes de María (Bolívar, Colombia), un macrofoco de leishmaniasis, se reportó el primer caso y se evidenciaron cambios en la distribución y ecoepidemiología de la enfermedad. No obstante, las interacciones entre vectores y fauna local aún son desconocidas. Objetivo. Evaluar la diversidad de flebotomíneos y sus fuentes de alimentación sanguínea en la comunidad de El Alférez del municipio de El Carmen de Bolívar (Bolívar, Colombia). Materiales y métodos. En el 2018, se recolectaron flebotomíneos mediante trampas de luz led ubicadas en el domicilio, el peridomicilio y en el área silvestre, y se identificaron a nivel de especie. Se utilizó la reacción en cadena de la polimerasa múltiple dirigida al gen mitocondrial citocromo B para analizar la sangre del aparato digestivo. Resultados. Lutzomyia evansi fue la especie más abundante (71,85 %; n = 485/675), seguida por Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci y Lu. aclydifera. El 25 % (n = 25/100) de las especies analizadas tuvieron como fuentes de ingesta sanguínea a Canis familiaris (36 %; n = 9/25), Ovis aries (36 %; n = 9/25), Bos taurus (24 %; n = 6/25), Sus scrofa (20 %; n = 5/25) y Homo sapiens (8 %; n = 2/25). Lutzomyia evansi fue la especie con la mayor frecuencia de alimentación (68 %; n = 17/25), predominantemente de una sola especie (44 %; n = 11/25) o de varias (24 %; n = 6/25).

Trypanosoma spp. infection in urban and wild ecotopes of the caribbean region in Colombia. / Infección por Trypanosoma spp. en murciélagos capturados en ecótopos urbanos y silvestres de la región Caribe en Colombia.

Motivation for the study. The role of bats as hosts of Trypanosoma spp. in the Atlantic department in Colombia, as well as its taxonomic diversity has been poorly studied. Main findings. This is the first report of frequency of infection by Trypanosoma spp. in bats in the Atlántico Department in Colombia. Implications. The great adaptive capacity of bats to different ecological niches and its role as hosts of Trypanosoma spp. for wild and urban ecotopes represents a risk factor in transmission cycles of epidemiological importance. We conducted a study to evaluate the frequency of infection by Trypanosoma spp. in bats captured in wild and urban ecotopes in the Department of Atlántico in the Caribbean region of Colombia from March 2021 to May 2022. Bats were taxonomically identified, and sex, relative age, and reproductive conditions were determined. A blood sample was used for parasitological analysis and DNA extraction to amplify a region of the 18S rRNA. 125 bats were collected, with the most abundant families being Molossidae (62/125; 49.6%) and Phyllostomidae (43/125; 34.4%). Molossus molossus collected in wild habitats showed an infection frequency of 8.1% (5/61) and 4.1% (3/61) through parasitological and molecular analysis, respectively. In comparison, Noctilio albiventris collected in urban habitats showed an infection frequency of 16.6% (2/12) for both analyses. These findings represent the first records of M. molossus harboring trypanosomes for the Department of Atlántico and of N. albiventris harboring trypanosomes in Colombia.

Se evaluó la frecuencia de infección por Trypanosoma spp. en murciélagos capturados en ecótopos silvestres y urbanos del Departamento del Atlántico, en la región Caribe de Colombia, entre marzo de 2021 y mayo de 2022. Se identificaron taxonómicamente los murciélagos y se determinó sexo, edad relativa y condiciones reproductivas. Se utilizó una muestra de sangre para análisis parasitológico y extracción de ADN para la amplificar una región del ARNr 18S. Se capturaron 125 murciélagos, siendo las familias más abundantes Molossidae (62/125; 49,6%) y Phyllostomidae (43/125; 34,4%). Molossus molossus capturado en ecótopos silvestres mostró una frecuencia de infección del 8,1% (5/61) y 4,1% (3/61) mediante análisis parasitológico y molecular, respectivamente. En comparación, Noctilio albiventris capturado en ecótopos urbanos mostró una frecuencia de infección del 16,6% (2/12) para ambos análisis. Estos hallazgos representan los primeros registros de M. molossus albergando Trypanosoma spp. para el Departamento del Atlántico y de N. albiventris albergando Trypanosoma spp. en Colombia. Motivación para realizar el estudio. El rol de los murciélagos como hospederos de Trypanosoma spp. en el Departamento del Atlántico en Colombia, así como su diversidad taxonómica ha sido poco estudiada. Principales hallazgos. Este es el primer reporte de frecuencia de infección por Trypanosoma spp. en murciélagos en el Departamento del Atlántico en Colombia. Implicancias. La gran capacidad de adaptación de los murciélagos a diferentes nichos ecológicos y su rol como hospederos de Trypanosoma spp. en ecótopos silvestres y urbanos representa un factor de riesgo en ciclos de transmisión de importancia epidemiológica.

Phlebotomine sand flies (Diptera: Psychodidae, Phlebotominae): Diversity of potential Leishmania vectors in northern Colombia.

Phlebotomine sand flies are critical vectors of Leishmania parasites, impacting public health significantly. This study focused on assessing the diversity of sand flies in a rural area of El Carmen de Bolívar Municipality, northern Colombia, employing rarefaction curves and Hill numbers to understand potential vector communities and inform environmental management. From January 2018 to April 2019 (five samplings), sand flies were collected using CDC light traps with blue LED in domestic/peridomestic/sylvatic ecotopes, identifying species per Young and Duncan (1994) and Galati (2003). Hill numbers provided diversity estimates across samples, while Principal Component Analysis correlated with environmental factors with phlebotomine species presence and abundance. 8,784 phlebotomine individuals were collected; 56.4 % females and 43.6% males (ratio 3:2). These individuals belonged to eight species: Pintomyia evansi, Psychodopygus panamensis, Lutzomyia gomezi, Micropygomyia cayennensis, Evandromyia dubitans, Psathyromyia aclydifera, Pintomyia serrana, and Pintomyia rangeliana; with Pi. evansi being the most abundant species (74.39 %; 6,530 exemplars). The ANOVA showed no significant differences between phlebotomine sand flies abundances across ecotopes (p = 0.018). Species of epidemiological relevance as Pi. evansi and Lu. gomezi not show a positive correlation with environmental variables evaluated, only Ps. panamensis was positively correlated with precipitation. However, the study emphasizes the need for a continuous sand fly monitoring and research to enhance leishmaniasis control strategies, highlighting the necessity to expand knowledge on phlebotomine diversity and environmental interactions to understand vector ecology and disease dynamics better.

Phlebotomine (Diptera: Psychodidae) species and their blood meal sources in a new leishmaniasis focus in Los Montes de María, Bolívar, in northern Colombia / Especies de flebotomíneos y sus fuentes de ingesta sanguínea en un nuevo foco de leishmaniasis en Los Montes de María (Bolívar) al norte de Colombia

Abstract Introduction. El Alférez, a village in Los Montes de María (Bolívar, Colombia) and a macro-focus of leishmaniasis, recorded its first case in 2018, evidencing changes in the distribution and eco-epidemiology of the disease, although interactions between vectors and local fauna remain unknown. Objective. To evaluate the diversity of sandflies and their blood meal sources in the community of El Alférez in the municipality of El Carmen de Bolívar (Bolívar, Colombia). Materials and methods. In 2018, sandflies were collected using LED-based light traps in domestic, peridomestic, and sylvatic ecotopes and identified at the species level. Multiplex polymerase chain reaction targeting the mitochondrial cytochrome B gene was used to analyze blood from the digestive tract. Results. Lutzomyia evansi was the most abundant species (71.85%; n = 485/675), followed by Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci, and Lu.aclydifera. Twenty-five percent of the species had blood meals from Canis familiaris (36.00%; n = 9/25), Ovis aries (36.00%; n=9:/25), Bos taurus (24.00%; n = 6/25), Sus scrofa (20.00%; n = 5/25), and Homo sapiens (8.00%; n = 2/25). Lutzomyia evansi registered the highest feeding frequency (68.00%; n = 17/25), predominantly on a single (44.00%; n = 11/25) or multiple species (24.00%; n = 6/25). Conclusion. Results indicate a eclectic feeding behavior in Lu. evansi, implying potential reservoir hosts for Leishmania spp. and increasing transmission risk. This study is a first step towards understanding the diversity of mammalian blood sources used by sandflies, that may be crucial for vector identification and formulation of effective control measures.

Resumen Introducción. En 2018, en la vereda El Alférez de Los Montes de María (Bolívar, Colombia), un macrofoco de leishmaniasis, se reportó el primer caso y se evidenciaron cambios en la distribución y ecoepidemiología de la enfermedad. No obstante, las interacciones entre vectores y fauna local aún son desconocidas. Objetivo. Evaluar la diversidad de flebotomíneos y sus fuentes de alimentación sanguínea en la comunidad de El Alférez del municipio de El Carmen de Bolívar (Bolívar, Colombia). Materiales y métodos. En el 2018, se recolectaron flebotomíneos mediante trampas de luz led ubicadas en el domicilio, el peridomicilio y en el área silvestre, y se identificaron a nivel de especie. Se utilizó la reacción en cadena de la polimerasa múltiple dirigida al gen mitocondrial citocromo B para analizar la sangre del aparato digestivo. Resultados. Lutzomyia evansi fue la especie más abundante (71,85 %; n = 485/675), seguida por Lu. panamensis, Lu. gomezi, Lu. trinidadensis, Lu. dubitans, Lu. abonnenci y Lu. aclydifera. El 25 % (n = 25/100) de las especies analizadas tuvieron como fuentes de ingesta sanguínea a Canis familiaris (36 %; n = 9/25), Ovis aries (36 %; n = 9/25), Bos taurus (24 %; n = 6/25), Sus scrofa (20 %; n = 5/25) y Homo sapiens (8 %; n = 2/25). Lutzomyia evansi fue la especie con la mayor frecuencia de alimentación (68 %; n = 17/25), predominantemente de una sola especie (44 %; n = 11/25) o de varias (24 %; n = 6/25). Conclusiones. Los hallazgos indican un comportamiento alimenticio ecléctico en Lu. evansi que implica potenciales reservorios para Leishmania spp. y eleva el riesgo de transmisión. Este estudio es un primer paso para comprender la diversidad de fuentes sanguíneas de mamíferos, utilizadas por los flebotomíneos, y que pueden ser cruciales para identificación de vectores y la formulación de medidas de control eficaces.

Chagas Disease and Leishmaniasis, Sympatric Zoonoses Present in Human from Rural Communities of Venezuela.

PURPOSE: Trypanosoma cruzi and Leishmania spp. coexist in several endemic areas, and there are few studies of Chagas disease and leishmaniasis coinfection worldwide; for this reason, the objective of this work was to determine the Chagas disease and leishmaniasis coinfection in several rural communities co-endemic for these diseases. METHODS: A total of 1107 human samples from six co-endemic rural communities of Cojedes state, Venezuela, were analyzed. Serum samples were evaluated by ELISA, indirect hemagglutination, and indirect immunofluorescence for Chagas disease diagnosis, and individuals were evaluated for leishmaniasis by leishmanin skin test (LST). Approximately, 30% of the individuals were also analyzed by PCR (blood clot samples) for T. cruzi and for Leishmania spp. RESULTS: The 14.7% of the individuals were positive to Trypanosoma cruzi infection by serology, and 25.8% were positive to Leishmania spp. current or past infection by LST. Among the group with PCR results, 7.8% were positive for T. cruzi, and 9.4% for Leishmania spp. The coinfection T. cruzi/Leishmania spp. was 6.5%. The T. cruzi DTUs of the positive blood clot samples were TcI, revealed using the molecular markers: (i) intergenic region of the miniexon, (ii) D7 divergent domain of the 24Sα rDNA, (iii) size-variable domain of the 18S rDNA, and (iv) hsp60-PCR-RFLP (EcoRV). The Leishmania species identified were L. (Leishmania) mexicana and L. (Viannia) braziliensis. CONCLUSION: A high prevalence was found for T. cruzi and Leishmania spp. single and coinfections in almost all communities studied, being these results of relevance for the implementation of control programs in co-endemic areas.

Infección por Trypanosoma spp. en murciélagos capturados en ecótopos urbanos y silvestres de la región Caribe en Colombia / Trypanosoma spp. infection in bats captured in urban and wild ecotopes of the Caribbean region in Colombia

R ESUMEN: Se evaluó la frecuencia de infección por Trypanosoma spp. en murciélagos capturados en ecótopos sil-vestres y urbanos del Departamento del Atlántico, en la región Caribe de Colombia, entre marzo de 2021 y mayo de 2022. Se identificaron taxonómicamente los murciélagos y se determinó sexo, edad relativa y condiciones reproductivas. Se utilizó una muestra de sangre para análisis parasitológico y extracción de ADN para la amplificar una región del ARNr 18S. Se capturaron 125 murciélagos, siendo las familias más abundantes Molossidae (62/125; 49,6%) y Phyllostomidae (43/125; 34,4%). Molossus molossus capturado en ecótopos silvestres mostró una frecuencia de infección del 8,1% (5/61) y 4,1% (3/61) mediante análisis parasitológico y molecular, respectivamente. En comparación, Noctilio albiventris capturado en ecótopos urbanos mostró una frecuencia de infección del 16,6% (2/12) para ambos análisis. Estos hallazgos re-presentan los primeros registros de M. molossus albergando Trypanosoma spp. para el Departamento del Atlántico y de N. albiventris albergando Trypanosoma spp. en Colombia.

Parasitological and molecular characterization of Trypanosoma cruzi of triatomines and mammals from endemic areas for Chagas disease in Venezuela.

It is estimated that 6-7 million people worldwide are infected with Trypanosoma cruzi, the parasite that causes Chagas disease. In Venezuela, Chagas disease remains a public health problem. In this work, T. cruzi isolates from six species of triatomines and mammals of the orders Didelphimorphia and Xenarthra, captured in rural communities of Monagas, underwent parasitological and molecular characterization. A total of 471 triatomines and 17 mammals were captured, with a natural infection rate of 41.4% and 70.6%, respectively. In the male NMRI mouse model used for parasitological characterization (prepatent period, parasitemia curve, mouse mortality, and tissular parasitism), T. cruzi isolates exhibited high lethality due to their pronounced virulence, irrespective of the parasite load in each mouse, resulting in a mortality rate of 75%. Among the vector isolates, in the mouse model, only 2 out of 6 remained alive, while the rest perished during the evaluation. Conversely, the isolates from mammals proved fatal for all the inoculated mice. All isolates were identified as belonging to DTU TcI, based on the molecular markers such as the intergenic region of the miniexon, D7 divergent domain of the 24Sα rDNA, size-variable domain of the 18S rDNA, and hsp60-PCR-RFLP-EcoRV. This study demonstrates the presence of vectors and mammalian reservoirs naturally infected with T. cruzi in communities of Monagas, the 9th largest and 9th most populous state in Venezuela. This situation represents a neglected epidemiological problem demanding urgent attention and imperative health care intervention.

Molecular detection of the natural infection by trypanosomatid parasites in Didelphis marsupialis from a rural area in northern Colombia. / Detección molecular de infección natural por parásitos tripanosomátidos en Didelphis marsupialis de una zona rural del norte de Colombia.

We studied the prevalence of infection by trypanosomatid parasites in Didelphis marsupialis and its relationship with morphological/age aspects in a rural area of El Carmen de Bolivar, Colombia. Five visits were made to the Vereda El Alférez; each of which lasted three consecutive nights. During these visits, Tomahawk® traps were installed in the peridomestic and wild ecotopes of the Vereda El Alférez. Body measurements, sex and age were determined from the collected animals. Blood was extracted by cardiopuncture, after sedation, in order to obtain total deoxyribonucleic acid (DNA) and amplify the conserved region of the kinetoplast minicircle DNA (kDNA) of parasitic trypanosomatids. The association between morphological parameters of didelphids and their frequency of infection by parasitic trypanosomatids was determined by binomial regression. Thirty D. marsupialis specimens (60.0% females and 40.0% males/66.7% adults and 33.3% juveniles) were collected. Molecular diagnosis revealed a frequency of trypanosomatid parasite infection of 46.7%. Stage (p=0.024) was a determinant for infection. We discuss the role of D. marsupialis as a potential reservoir of parasitic trypanosomatids in the Vereda El Alférez.

Se evaluó la prevalencia de infección por parásitos tripanosomátidos en Didelphis marsupialis y su relación con los aspectos morfológicos/etarios en una zona rural de El Carmen de Bolívar, Colombia. En cinco visitas (2018-2019) se instalaron trampas Tomahawk® en los ecótopos peridoméstico y silvestre en la Vereda El Alférez, durante tres noches consecutivas/visita. A los animales recolectados, se les determinaron medidas corporales, sexo y edad; y se les extrajo sangre por cardiopuntura, previa sedación, para extracción del ácido desoxirribonucleico (ADN) total y amplificación de la región conservada del ADN de minicírculos de kinetoplasto (ADNk) de parásitos tripanosomátidos. La asociación entre parámetros morfológicos de los didélfidos y su frecuencia de infección por parásitos tripanosomátidos fue determinada mediante una regresión binomial. Se recolectaron 30 individuos de D. marsupialis (60,0% hembras y 40,0% machos/66,7% adultos y 33,3% juveniles). El diagnóstico molecular reveló una frecuencia de infección por parásitos tripanosomátidos del 46,7%. El estadio (p=0,024) fue determinante para la infección. Se discute el papel de D. marsupialis como potencial reservorio de parásitos tripanosomátidos en la zona evaluada.

Geographic Distribution of the Genus Panstrongylus Berg, 1879 in the Neotropic with Emphasis on Trypanosoma cruzi Vectors.

Panstrongylus is a Neotropical taxa of 16 species, some more widespread than others, that act as vectors of Trypanosoma cruzi, the etiologic agent of Chagas disease (CD). This group is associated with mammalian reservoir niches. There are few studies of the biogeography and niche suitability of these triatomines. Using zoo-epidemiological occurrence databases, the distribution of Panstrongylus was determined based on bioclimatic modelling (DIVA GIS), parsimonious niche distribution (MAXENT), and parsimony analysis of endemic species (PAE). Through 517 records, a wide presence of P. geniculatus, P. rufotuberculatus, P. lignarius, and P. megistus was determined and recorded as frequent vectors of T. cruzi in rainforest habitats of 24-30 °C. These distributions were modeled with AUC >0.80 and <0.90, as well as with the seasonality of temperature, isothermality, and precipitation as relevant bioclimatic variables. Individual traces for each taxon in Panstrongylus-1036 records-showed widely dispersed lines for frequent vectors P. geniculatus, P. lignarius, P. rufotuberculatus, and P. megistus. Other occasional vectors showed more restricted dispersal, such as P. howardi, P. humeralis, P. lenti, P. lutzi, P. tupynambai, P. noireaiui, and P. chinai. Areas of defined environmental variation, geological change, and trans domain fluid fauna, such as the American Transition Zone and the Pacific Domain of Morrone, had the highest Panstrongylus diversity. Pan-biogeographic nodes appear to be areas of the greatest species diversity that act as corridors connecting biotopes and allowing fauna migration. Vicariance events in the geologic history of the continent need to be investigated. The geographical distribution of Panstrongylus overlapped with CD cases and Didelphis marsupialis/Dasypus novemcinctus presence, two important reservoirs in Central and South America. The information derived from the distribution of Panstrongylus provides knowledge for surveillance and vector control programs. It would increase information on the most and less relevant vector species of this zoonotic agent, for monitoring their population behavior.

Detección molecular de la infección natural por parásitos tripanosomátidos en Didelphis marsupialis de una zona rural del norte de Colombia / Molecular detection of the natural infection by trypanosomatid parasites in Didelphis marsupialis from a rural area in northern Colombia

Se evaluó la prevalencia de infección por parásitos tripanosomátidos en Didelphis marsupialis y su relación con los aspectos morfológicos/etarios en una zona rural de El Carmen de Bolívar, Colombia. En cinco visitas (2018-2019) se instalaron trampas Tomahawk® en los ecótopos peridoméstico y silvestre en la Vereda El Alférez, durante tres noches consecutivas/visita. A los animales recolectados, se les determinaron medidas corporales, sexo y edad; y se les extrajo sangre por cardiopuntura, previa sedación, para extracción del ácido desoxirribonucleico (ADN) total y amplificación de la región conservada del ADN de minicírculos de kinetoplasto (ADNk) de parásitos tripanosomátidos. La asociación entre parámetros morfológicos de los didélfidos y su frecuencia de infección por parásitos tripanosomátidos fue determinada mediante una regresión binomial. Se recolectaron 30 individuos de D. marsupialis (60,0% hembras y 40,0% machos/66,7% adultos y 33,3% juveniles). El diagnóstico molecular reveló una frecuencia de infección por parásitos tripanosomátidos del 46,7%. El estadio (p=0,024) fue determinante para la infección. Se discute el papel de D. marsupialis como potencial reservorio de parásitos tripanosomátidos en la zona evaluada.

We studied the prevalence of infection by trypanosomatid parasites in Didelphis marsupialis and its relationship with morphological/age aspects in a rural area of El Carmen de Bolivar, Colombia. Five visits were made to the Vereda El Alférez; each of which lasted three consecutive nights. During these visits, Tomahawk® traps were installed in the peridomestic and wild ecotopes of the Vereda El Alférez. Body measurements, sex and age were determined from the collected animals. Blood was extracted by cardiopuncture, after sedation, in order to obtain total deoxyribonucleic acid (DNA) and amplify the conserved region of the kinetoplast minicircle DNA (kDNA) of parasitic trypanosomatids. The association between morphological parameters of didelphids and their frequency of infection by parasitic trypanosomatids was determined by binomial regression. Thirty D. marsupialis specimens (60.0% females and 40.0% males/66.7% adults and 33.3% juveniles) were collected. Molecular diagnosis revealed a frequency of trypanosomatid parasite infection of 46.7%. Stage (p=0.024) was a determinant for infection. We discuss the role of D. marsupialis as a potential reservoir of parasitic trypanosomatids in the Vereda El Alférez.

Genetic diversity of Aedes aegypti in the central-upper Paraná Cross-Border axis in Paraguay. / Diversidad genética de aedes aegypti en el eje transfronterizo Central-Alto Paraná en Paraguay.

OBJECTIVE: To determine the genetic diversity of Aedes aegypti in the Central-Alto Paraná cross-border road corridor of Paraguay, an area that has reports of dengue cases. MATERIALS AND METHODS: Twenty adult females were selected from hatching Ae. aegypti eggs from households geolocated in the departments of Alto Paraná, Caaguazú, Cordillera and Central, between 2018 and 2019. DNA was extracted from the tissue of females for amplifying their polymorphic patterns by random amplification of polymorphic DNA by PCR (RAPD-PCR), using primers H3 and B03 in order to identify genetic parameters of population diversity. The relationships between mosquito populations according to locality were observed by unpaired arithmetic mean analysis. We used DIVA-GIS 7.3.0 and MAXENT to analyze the suitable areas of actual and potential geographic distribution of these Ae. aegypti populations. RESULTS: Forty loci were identified by RAPD-PCR profiling, with moderate gene differentiation (Gst = 0.12). The cross-border corridor presented bioclimatic conditions for the presence of variant populations of Ae. aegypti, with precipitation in the warmest quarter and mean temperature in the driest quarter being determinant in the distribution. CONCLUSIONS: There is evidence of moderate genetic diversity in Ae. aegypti populations from areas that have reported dengue cases in the cross-border road corridor linking the Central and Alto Paraná departments of Paraguay. The study of genetic variability of Ae. aegypti is very useful for entomo-epidemiological surveillance and evaluation of possible resistance to chemical control.

OBJETIVOS: Conocer la diversidad genética de Aedes aegypti en el corredor vial transfronterizo Central-Alto Paraná de Paraguay, con registros de casos de dengue. MATERIALES Y MÉTODOS: Se seleccionaron veinte hembras adultas de la eclosión de huevos de Ae. aegypti procedentes de casas geolocalizadas en los departamentos de Alto Paraná, Caaguazú, Cordillera y Central, entre el 2018 y 2019. Se extrajo ADN del tejido de las hembras para amplificación aleatoria de sus patrones polimórficos mediante amplificación aleatoria del ADN polimórfico por PCR (RAPD-PCR), usando cebadores H3 y B03 a fin de conocer parámetros genéticos de diversidad poblacional. Las relaciones entre las poblaciones de mosquitos según la localidad fueron visualizadas mediante análisis no apareado de la media aritmética. Las áreas idóneas de distribución geográfica real y potencial de estas poblaciones de Ae. aegypti fueron analizadas mediante DIVA-GIS 7.3.0 y MAXENT. RESULTADOS: Se identificaron 40 loci mediante perfiles RAPD-PCR, con diferenciación génica moderada (Gst = 0,12). El corredor transfronterizo presentó condiciones bioclimáticas para la presencia de poblaciones variantes de Ae. aegypti, siendo determinantes en la distribución la precipitación del trimestre más cálido y la temperatura media del trimestre más seco. CONCLUSIONES: Se evidencia que existe diversidad genética moderada en las poblaciones de Ae. aegypti procedentes de zonas con registros de casos de dengue ubicadas en el corredor vial transfronterizo que une los departamentos Central y Alto Paraná de Paraguay. El estudio de variabilidad genética de Ae. aegypti es de gran utilidad para la vigilancia entomoepidemiológica y evaluación de posibles eventos de resistencia al control químico.

FLEBOTOMINOFAUNA Y PRIMER REGISTRO DE Lutzomyia aclydifera (DIPTERA: PSYCHODIDAE) EN EL CARIBE COLOMBIANO / Phlebotomine fauna and the first record of Lutzomyia aclydifera (Díptera: Psychodidae) in the Colombian Caribbean

RESUMEN Uno de los focos de leishmaniasis más importante de Colombia está en Los Montes de María, Departamento de Bolívar, específicamente en El Carmen de Bolívar. A la fecha están registradas 19 especies del género Lutzomyia, en este departamento, algunas con antecedentes como vectores de Leishmania spp. La presente nota informa la abundancia y la diversidad de la flebotominofauna en la zona rural de El Carmen de Bolívar. Mediante el uso de trampas CDC, se colectaron 1327 hembras del género Lutzomyia entre enero a noviembre de 2018: Lu. evansi (81 %), Lu. panamensis (15,5 %), Lu. gomezi (2,9 %), Lu. dubitans (0,15 %), Lu. aclydifera (0,07 %), Lu. c. cayennensis (0,07 %), Lu. rangeliana (0,07 %) y Lu. serrana (0,07 %). La mayor abundancia de flebotomíneos se presentó en noviembre y la menor en abril, coincidiendo con los meses de lluvia y sequía, respectivamente; mientras que en enero se presentó la mayor diversidad. En esta investigación sobresalen Lu. evansi, Lu. panamensis y Lu. gomezi por sus antecedentes como vectores de Leishmania en el Departamento de Bolívar y otras regiones del país. Se reporta por primera vez para el Caribe Colombiano la especie Lu. aclydifera, la cual deberá ser vigilada en cuanto a su capacidad vectorial para Leishmania.

ABSTRACT One of the most important leishmaniasis foci in Colombia is Los Montes de María, Department of Bolívar, specifically in El Carmen de Bolívar. To date, 19 species of Lutzomyia have been recognized in this department, some with antecedents as vectors of Leishmania spp. This note reports the abundance and diversity of the phlebotomine sandflies fauna in the rural area of El Carmen de Bolívar. Through the use of CDC traps, 1327 females of the genus Lutzomyia were collected between January to November 2018: Lu. evansi (81 %), Lu. panamensis (15.5 %), Lu. gomezi (2.9 %), Lu. dubitans (0.15 %), Lu. aclydifera (0.07 %), Lu. c. cayennensis (0.07 %), Lu. rangeliana (0.07 %) and Lu. serrana (0.07 %). The highest abundance of sandflies occurred in November and the lowest in April, which is consistent with the rainy and dry months, respectively; while in January there was the greatest diversity. In this research, stands out Lu. evansi, Lu. panamensis and Lu. gomezi for their antecedents as vectors of Leishmania in the Department of Bolívar and other regions of the country. We reported the species Lu. aclydifera for the first time for the Caribbean region, hence, this species should be monitored given its vector capacity for Leishmania.

Molecular diagnosis of trypanosomatids in Didelphis marsupialis from Los Montes de María: a first report of Trypanosoma rangeli from Colombian Caribbean region.

Didelphis marsupialis is a primary reservoir of Trypanosoma cruzi, etiologic agent of American Trypanosomiasis-AT or Chagas Disease-CD, in America. Some findings of Trypanosoma rangeli have been recorded in this mammal, in sympatry with T. cruzi. In Los Montes de María, Bolívar, Colombian Caribbean, triatomine insects and potential parasite host has been registered, but little is known about the relationship between these parasites and D. marsupialis. We investigated the natural trypanosomatids infection rate in D. marsupialis, applying a parasitological and molecular diagnosis. Twenty D. marsupialis was investigated between 2018 and 2019 using 21 Tomahawk® traps placed on the sylvatic/domestic corridors. Blood was drawn by cardiopuncture after sedation. An aliquot of blood samples was cultured in Novy, Nicolle, McNeal/Roswell Park Memorial Institute medium at 24 °C/60 days for the detection of motile trypomastigotes. Parasite DNA was obtained by salting out methods from positive blood cultures. Trypanosomatids diagnosis was done by Polymerase Chain Reaction-sequencing of V7V8 region of 18S ribosomal RNA (18S-rRNA) gene. Amplicons were sequenced, and consensus sequences were aligned with reference sequences from GenBank. Four isolates corresponded to T. rangeli (20%) and one to T. cruzi (5%). The natural infection of D. marsupialis by T. rangeli and T. cruzi constitutes the first record of these parasites in didelphids in Los Montes de María and the first record of T. rangeli in this marsupial, in the Colombian Caribbean.

Diversidad genética de aedes aegypti en el eje transfronterizo Central-Alto Paraná en Paraguay / Genetic diversity of Aedes aegypti in the central-upper Paraná Cross-Border axis in Paraguay

RESUMEN Objetivos: Conocer la diversidad genética de Aedes aegypti en el corredor vial transfronterizo Central-Alto Paraná de Paraguay, con registros de casos de dengue. Materiales y métodos: Se seleccionaron veinte hembras adultas de la eclosión de huevos de Ae. aegypti procedentes de casas geolocalizadas en los departamentos de Alto Paraná, Caaguazú, Cordillera y Central, entre el 2018 y 2019. Se extrajo ADN del tejido de las hembras para amplificación aleatoria de sus patrones polimórficos mediante amplificación aleatoria del ADN polimórfico por PCR (RAPD-PCR), usando cebadores H3 y B03 a fin de conocer parámetros genéticos de diversidad poblacional. Las relaciones entre las poblaciones de mosquitos según la localidad fueron visualizadas mediante análisis no apareado de la media aritmética. Las áreas idóneas de distribución geográfica real y potencial de estas poblaciones de Ae. aegypti fueron analizadas mediante DIVA-GIS 7.3.0 y MAXENT. Resultados: Se identificaron 40 loci mediante perfiles RAPD-PCR, con diferenciación génica moderada (Gst = 0,12). El corredor transfronterizo presentó condiciones bioclimáticas para la presencia de poblaciones variantes de Ae. aegypti, siendo determinantes en la distribución la precipitación del trimestre más cálido y la temperatura media del trimestre más seco. Conclusiones: Se evidencia que existe diversidad genética moderada en las poblaciones de Ae. aegypti procedentes de zonas con registros de casos de dengue ubicadas en el corredor vial transfronterizo que une los departamentos Central y Alto Paraná de Paraguay. El estudio de variabilidad genética de Ae. aegypti es de gran utilidad para la vigilancia entomoepidemiológica y evaluación de posibles eventos de resistencia al control químico.

ABSTRACT Objective: To determine the genetic diversity of Aedes aegypti in the Central-Alto Paraná cross-border road corridor of Paraguay, an area that has reports of dengue cases. Materials and methods: Twenty adult females were selected from hatching Ae. aegypti eggs from households geolocated in the departments of Alto Paraná, Caaguazú, Cordillera and Central, between 2018 and 2019. DNA was extracted from the tissue of females for amplifying their polymorphic patterns by random amplification of polymorphic DNA by PCR (RAPD-PCR), using primers H3 and B03 in order to identify genetic parameters of population diversity. The relationships between mosquito populations according to locality were observed by unpaired arithmetic mean analysis. We used DIVA-GIS 7.3.0 and MAXENT to analyze the suitable areas of actual and potential geographic distribution of these Ae. aegypti populations. Results: Forty loci were identified by RAPD-PCR profiling, with moderate gene differentiation (Gst = 0.12). The cross-border corridor presented bioclimatic conditions for the presence of variant populations of Ae. aegypti, with precipitation in the warmest quarter and mean temperature in the driest quarter being determinant in the distribution. Conclusions: There is evidence of moderate genetic diversity in Ae. aegypti populations from areas that have reported dengue cases in the cross-border road corridor linking the Central and Alto Paraná departments of Paraguay. The study of genetic variability of Ae. aegypti is very useful for entomo-epidemiological surveillance and evaluation of possible resistance to chemical control.

Susceptibilidad a deltametrina de Aedes (Stegomyia) aegypti (Linnaeus 1676) (Diptera: Culicidae) y determinación de áreas de influencia para su presencia en un municipio del Departamento Central, Paraguay / Susceptibility to deltametrin of Aedes (Stegomyia) aegypti(Linnaeus 1676) (Diptera: Culicidae) and determination from areas of influence for its presence in a municipality of the Centraldepartment, Paraguay

RESUMEN Introducción. Paraguay es un país endémico para el dengue y la intervención primaria para esta arbovirosis es el control químico de su vector Aedes aegypti, siendo necesario estudiar su resistencia frente a insecticidas utilizados para el control y conocer su sostenibilidad en espacio y tiempo. Objetivo. Determinar la susceptibilidad de poblaciones silvestres de Ae. aegypti procedentes de Villa Elisa-Central, frente al adulticida deltametrina y conocer el área potencial de distribución de estas poblacionesresistentes/susceptibles. Metodología. Se utilizaron mosquitos Ae. aegypti adultos de la Filial 1 obtenidos de ovitrampas colocadas en viviendas geo-localizadas entre diciembre 2017-marzo 2018. Las hembras resultantes de la cría en condiciones ad hoc en laboratorio fueron expuestas a concentraciones crecientes del insecticida. La distribución real de las poblaciones de mosquitos y su área de influencia, fueron modeladas mediante DIVA-GIS. Resultados. La mortalidad promedio observada para cada dosis de deltametrina fue: 5,3% para (0,03%); 47,4% para (0,15%) y 61,2% para (0,3). Se observó áreas de influencia para la presencia de Ae. aegypti. Conclusión. los valores correspondientes a la mortalidad resultaron inferiores al 98%, lo cual fue indicativo de resistencia a dosis diagnóstica. El modelo de distribución geográfica de la población evaluada demostró la existencia de áreas idóneas con condiciones bioclimáticas óptimas (niveles de precipitación, temperatura y humedad), propicias para la aparición de reservorios de agua y presencia del vector, tanto en Villa Elisa, como en los distritos localizados en la periferia tales como San Lorenzo, Lambaré, Ñemby, San Antonio y Fernando de la Mora.

ABSTRACT Introduction. Paraguay is an endemic country for dengue and the primary intervention for this arbovirus is the chemical control of its vector Aedes aegypti, being necessary to study its resistance against insecticides used for control and to know its sustainability in space and time. Objective. To determine the susceptibility of wild populations of Ae. aegypti from Villa Elisa-Central, against the adulticide deltamethrin and to know the potential area of ​​distribution of these resistant/susceptible populations. Methodology. Mosquitoes Ae. aegypti adults from Filial 1 obtained from ovitraps placed in geo-located dwellings between December 2017 and March 2018. The females resulting from rearing under ad hoc conditions in the laboratory were exposed to increasing concentrations of the insecticide. The actual distribution of mosquito populations and their area of ​​influence were modeled using DIVA-GIS. Results. The average mortality observed for each dose of deltamethrin was: 5.3% for (0.03%); 47.4% for (0.15%) and 61.2% for (0.3). Areas of influence were observed for the presence of Ae. aegypti. Conclusion. The values ​​corresponding to mortality were lower than 98%, which was indicative of resistance to diagnostic dose. The geographic distribution model of the evaluated population demonstrated the existence of suitable areas with optimal bioclimatic conditions (levels of precipitation, temperature and humidity), conducive to the appearance of water reservoirs and the presence of the vector, both in Villa Elisa and in the districts located on the outskirts such as San Lorenzo, Lambaré, Ñemby, San Antonio and Fernando de la Mora.

Infections and Coinfections by Trypanosomatid Parasites in a Rural Community of Venezuela.

INTRODUCTION: Trypanosoma cruzi, Trypanosoma rangeli and Leishmania spp. are parasites that coexist in several endemic areas. The identification of these parasites in hosts is important for the control programs. METHODS: 216 samples from human blood (101), blood of other mammals (45) and triatomine intestinal content and hemolymph (70), from an endemic area of Venezuela, were analysed. The samples were evaluated by; serology (only humans) and PCR for T. cruzi in human, other mammals and triatomines, PCR for T. rangeli in mammals-including human and triatomines and PCR for Leishmania in mammals-including human. RESULTS: The 9.9% of the human samples were positive for T. cruzi by serology, 11.9% by PCR, 4% for T. rangeli PCR and none for Leishmania spp. PCR. 60% of the samples of other mammals showed DNA amplification for T. cruzi, 42.2% for T. rangeli and 4.4% for Leishmania spp. 61.4% of the triatomine samples showed DNA amplification for T. cruzi and 10% for T. rangeli. CONCLUSIONS: High T. cruzi infection was detected in mammals and triatomines compared with T. rangeli. Low leishmanial infection was detected in other mammals. It is the first time that T. cruzi/T. rangeli coinfection, in humans, Canis familiaris (dog), and Bos Taurus (cow), were reported world-wide, and that this coinfection was described in Tamandua tetradactyla (anteater) from Venezuela. The coinfection T. cruzi/T. rangeli in mammals-including humans and triatomines, and coinfection T. cruzi/Leishmania spp. in non-human mammals, show the risk for trypanosomic zoonoses in this endemic area.

Evaluation of commercial kits for the immunological and molecular diagnosis of Chagas disease in endemic areas of Venezuela.

INTRODUCTION: The sensitivity and specificity of diagnostic techniques for Chagas disease depend largely on the antigens and targets used and on the immune response and characteristics of the infection of the population where it is applied, hence the need for evaluation of the diagnostic techniques available in a given area. So, the objective of this work was to evaluate two commercial kits for the immunological and molecular diagnosis of Chagas disease in endemic areas of Venezuela. METHODS: The evaluated kits were: Chagas ELISA IgG+IgM® and Speed Oligo Chagas® (Vircell®, Granada, Spain). They were evaluated with 129 samples (35 from patients in the acute phase, 33 in the chronic phase, 31 from patients with other diseases, and 30 from healthy individuals). The results were compared with those obtained in the conventional ELISA and PCR-satellite DNA tests for Trypanosoma cruzi. RESULTS: With Chagas ELISA IgG+IgM® a sensitivity of 94.1% and specificity of 93.4% were obtained, with Speed Oligo Chagas® a sensitivity of 92.6% and specificity of 100% were achieved, values similar to those showed by conventional ELISA and satDNA-PCR. CONCLUSION: The sensitivity and specificity of the commercial kits evaluated make them suitable for the diagnosis of Chagas disease in endemic areas of Venezuela.

Manifestaciones oculares de la toxocariasis en escolares del estado Anzoátegui en Venezuela / Ocular manifestations of toxocariasis in schoolchildren from the state from the Anzoátegui state in Venezuela

RESUMEN El objetivo de este estudio fue determinar manifestaciones oculares de la toxocariasis en escolares. Se realizó un estudio en dos escuelas del estado Anzoátegui en Venezuela en el 2019. Se empleó la prueba de ELISA para determinar los anticuerpos IgG contra Toxocara spp. Las familias completaron un cuestionario y los niños fueron evaluados clínicamente por pediatras y oftalmólogos. Participaron 118 niños, el 18,6% presentó anticuerpos anti-Toxocara spp. Las manifestaciones clínicas con asociación estadísticamente significativa fueron las reacciones alérgicas, epífora y disminución de la agudeza visual. En la evaluación oftalmológica se encontró queratitis, uveítis, iritis, granuloma retiniano, endoftalmitis, amaurosis, leucocoria, desprendimiento de retina y endotropía. Los hallazgos muestran una alta frecuencia de enfermedad ocular en niños con toxocariasis de un estado de Venezuela.

ABSTRACT The objective of this study was to determine ocular manifestations of toxocariasis in schoolchildren. A study was conducted in two schools in the Anzoátegui state in Venezuela in 2019. The ELISA test was used to determine IgG antibodies against Toxocara spp. The families completed a questionnaire, and the children were clinically evaluated by pediatricians and ophthalmologists. 118 children participated, 18.6% presented anti-Toxocara spp. The clinical manifestations with a statistically significant association were allergic reactions, epiphora, and decreased visual acuity. The ophthalmological evaluation found keratitis, uveitis, iritis, retinal granuloma, endophthalmitis, amaurosis, leukocoria, retinal detachment and endotropia. The findings show a high frequency of eye disease in children with toxocariasis from a state of Venezuela.

Ecopathogenic complexes of American trypanosomiasis in endemic areas of Venezuela: Diagnosis and variability of Trypanosoma cruzi.

BACKGROUND & OBJECTIVES: Trypanosoma cruzi, the causative agent of American trypanosomiasis, has been reported in 180 mammalian species and 154 triatomines species of Neotropic. This is a clonal parasite with variable biological behaviour, associated with the genetics of the parasite and its hosts. To know the eco-pathogenic complex of this zoonosis, it was proposed to characterize T. cruzi isolates obtained from triatomines and domestic, peridomestic and wild mammals of the eastern and central-western regions of Venezuela. METHODS: The positivity to T. cruzi was established and the isolates were genetically characterized by PCR amplification of the mini-exon gene, the DNA coding for 24Sa and 18S rRNA, and polymorphic sequences-RFLPs. The sampling sites were georeferenced using the MapSource Software and ArcGis 9.3 programs to generate distribution maps of the isolates. RESULTS: Of the 460 hosts (205 triatomines and 255 mammals), 49% were positive for the parasite. On the other hand, 38 isolates obtained from the triatomines and 23 isolates obtained from mammals were evaluated. The TcI genotype predominated in most of the isolates; however, in those obtained from triatomines the presence of the TcIII genotype in single infections and TcI + TcIII or TcI + TcIV in mixed infections was also evidenced. INTERPRETATION & CONCLUSION: There is a possibility that the triatomines act as biological syringes for these genotypes associated exclusively to them. The heterogeneity in T. cruzi isolates demonstrated the complexity of parasitosis in these regions, presenting its control and prevention as a challenge.

First molecular detection of Trypanosoma cruzi, T. rangeli and Leishmania spp. in capybaras.

Hydrochoerus hydrochaeris (capybara), is a widely distributed rodent in Latin America, with exploitation for food purposes and also used in leather industry products. The infection of this rodent by trypanosomatids may not be detected by parasitological methods, due to low parasitemias. The Capybaras blood samples from the Apure State were collected on filter paper, DNA was extracted and PCR was performed. The PCR technique was used for the detection of Trypanosoma cruzi satellite and kinetoplast DNA, T. rangeli miniexon, T. evansi RIME sequence, and DNA encoding ribosomal RNA and internal transcribed spacer 1 from Leishmania spp. Of the 16 evaluated samples, 12 (75%) were positive for T. cruzi, two for T. rangeli (12.5%), one for Leishmania spp. (6.3%) and none for T. evansi. Regarding coinfection, the two specimens infected with T. rangeli were also infected with T. cruzi (12.5%) and the positive sample for Leishmania spp. was also infected with T. cruzi (6.3%). The results shown in this study represent the first finding of T. cruzi infection, detected by molecular methods, world-wide and the first time that T. rangeli and Leishmania spp. have been found in capybaras. In addition, we report coinfections by T. cruzi/T. rangeli and T. cruzi/Leishmania spp. in H. hydrochaeris for the first time world-wide. Capybaras are widely managed as a source of animal protein, the results obtained require evaluating their possible role as a reservoir in trypanosomiasis and leishmaniasis. A 'One Health' approach through combination of ecological, veterinary and human health including the diagnosis and treatment of diseases of both humans and animals is essential for the development of more successful health programs.