Rhabdoviral Endogenous Sequences Identified in the Leishmaniasis Vector Lutzomyia longipalpis Are Widespread in Sandflies from South America.

Sandflies are known vectors of leishmaniasis. In the Old World, sandflies are also vectors of viruses while little is known about the capacity of New World insects to transmit viruses to humans. Here, we relate the identification of RNA sequences with homology to rhabdovirus nucleocapsids (NcPs) genes, initially in the Lutzomyia longipalpis LL5 cell lineage, named NcP1.1 and NcP2. The Rhabdoviridae family never retrotranscribes its RNA genome to DNA. The sequences here described were identified in cDNA and DNA from LL-5 cells and in adult insects indicating that they are transcribed endogenous viral elements (EVEs). The presence of NcP1.1 and NcP2 in the L. longipalpis genome was confirmed in silico. In addition to showing the genomic location of NcP1.1 and NcP2, we identified another rhabdoviral insertion named NcP1.2. Analysis of small RNA molecules derived from these sequences showed that NcP1.1 and NcP1.2 present a profile consistent with elements targeted by primary piRNAs, while NcP2 was restricted to the degradation profile. The presence of NcP1.1 and NcP2 was investigated in sandfly populations from South America and the Old World. These EVEs are shared by different sandfly populations in South America while none of the Old World species studied presented the insertions.

Nyssomyia whitmani (Diptera: Psychodidae) Activity From Sunset to Sunrise: A Two-Year Study in a Rural Area in Northeast Argentina.

We model the nocturnal patterns of the main vector of Leishmania braziliensis (Vianna) in the Paranaense region of South America, Nyssomyia whitmani (Antunes & Coutinho). Occurrence and abundance variation were analyzed by hourly periods, and in relation to sunrise and sunset to avoid differences in luminosity at fixed times throughout the year. The possible role of climatic and environmental variables on nocturnal activity curves was explored. A rotating light trap was operated biweekly in the peridomestic henhouse of a farm in Puerto Iguazú, from May 2016 to May 2018. Ny. whitmani, is the predominant Phlebotominae species (6,720 specimens captured), and is present throughout the night and seasons. Generalized additive models of nocturnal abundance of Ny. whitmani show a bell shape with an acrophase in the early evening. Models of abundance distribution as a function of total abundance showed significant nonlinear variations, only for the tertile of highest vector abundance, in the curves by hourly periods, and related to dawn, while female abundance relative to total abundance showed an asymptotic increasing curve, suggesting density-dependent events with abundance thresholds. Finally, temperature was the variable that best explained the pattern of abundance distribution of Ny. whitmani during the night, although triggers for scotophase activity due to internal clocks or luminosity could not be ruled out. The relationship of activity and abundance with climatic variables would also explain the nights of extraordinary abundance, and therefore of greater probability of vector-human and vector-reservoir contacts.

The second coming of urban yellow fever in the Americas: looking the past to see the future.

Yellow fever (YF) epizootics in South America during the 21st century have an unprecedented recorded magnitude and geographical dispersion. The YF spread progressively involved areas considered previously free of risk reaching the edge of cities with large unvaccinated populations, and urban outbreaks were frequently forecasted. We critically reviewed the initial stages and enhancing contexts of YF urban epidemics since the 17th century in the Americas, and the modeling attempts of YF epidemic risk by of Aedes-Human transmission, to find common factors that increase the probability of these events in the current scenarios. The YF urban outbreaks of the past showed as necessary conditions the multiple introduction by viremic carriers clustered in time and space, coincident with population peaks of Aedes. These conditions are not met in the current outbreaks in the Americas by sylvatic YF cycles, besides the protective impact of vector control campaigns, vaccination coverage, improved surveillance, and case management. Therefore, urban Aedes-Human YF outbreaks in the Americas are still possible but with low probability or very focal transmission, while the conditions reported in the past were avoided, and the surveillance and control measures sustained, including the vaccination of the population at risk.

Visceral Leishmaniasis in Bolivia: Current Status.

INTRODUCTION: In Bolivia, before 1982 there were no records of visceral leishmaniasis (VL) cases that would allow us to review and describe the temporospatial occurrence of VL by ecoregions in provinces and departments of Bolivia to evaluate its impact on public health, risk of outbreaks, or dispersion. METHODS: This update on VL in Bolivia is based on research, reviews, and retrospective literature analyses of online data and libraries and institutional reports, from 1939 to the present. RESULTS: In Bolivia, 56 cases of VL have been reported. Until 2014, only three endemic departments had been identified (La Paz, Santa Cruz, and Tarija). Since then, further cases have been recorded in Pando, Cochabamba, and Beni, and in Chuquisaca in 2015. In Yungas, a VL focus was confirmed by isolating and comparing parasites from human and dog cases, and from the Lu. longipalpis vector. VL cases from seven departments, involving 12 different ecoregions were located within the Amazon and Plata basins. CONCLUSIONS: We confirmed that dogs are its primary reservoir, and Lutzomyia longipalpis is its main vector (currently dispersed in six departments). The primary vectors in areas where Lutzomyia longipalpis is absent are Migonemyia migonei and Lutzomyia cruzi.

Niche divergence and paleo-distributions of Lutzomyia longipalpis mitochondrial haplogroups (Diptera: Psychodidae).

Lutzomyia longipalpis is a complex of species which has a wide but discontinuous distribution from southeastern Mexico to northern Argentina and Uruguay. To date, eight mitochondrial haplogroups have been identified along its distribution although key environmental tolerances and ecological niche models have been analyzed only at the complex level. The aim of the present study was to analyze whether genetic diversification using three mitochondrial genes of the Lu. longipalpis complex is associated with niche divergence and to explore evolution of distributional projections of all haplogroups between the Last Glacial Maximum (LGM; 21,000 yrs ago) and the present. Current occurrence of all haplogroups was used to develop ecological niche models (ENM) and these were projected in both periods to quantify and identify geographic area shifts. Environmental space was used to estimate niche similarity between major clades and pairwise between individual haplogroups. The two major Lu. longipalpis clades (Mex, CA, Col and Ven vs Arg and Bra) had significantly different environmental space, indicating niche divergence. Environmental space overlap of southern haplogroups was variable, with divergent niche, except between Arg and ArgBra. The most suitable regions for the ArgBra haplogroup were northeastern and southeastern Brazil, and the Gran Chaco region. In contrast, ENM of haplogroups within the northern major clade have significantly similar niche, with highest geographic ENM suitability along both the Caribbean and Pacific coasts. The intensity and coverage of high suitability areas in the LGM decreased for most haplogroups in the present. Integrating ENM and phylogenetic analyses has allowed us to test hypotheses of niche similarity between Lu. longipalpis haplogroups and major clades, and to identify conserved distributional areas of haplogroups since the LGM, with the exception of Arg. Evidence for distributional shifts and overlap of haplogroups is important to analyze Leishmaniasis´ eco-epidemiology and to successfully monitor and control transmission.

Leishmaniasis en Bolivia. Comprehensive review and current status in Tarija, in the border with Argentina / [Leishmaniasis en Bolivia, revisión y estado actual en Tarija, frontera con Argentina].

Introduction: In the department of Tarija in the Bolivian-Argentine border, human cases with ulcers on uncovered parts of the skin plus clinical and epidemiological characteristics related to leishmaniasis were reported for the first time in 1997. Objective: To describe and to verify the presence of leishmaniasis in Tarija, sixth endemic department in Bolivia. Materials and methods: We conducted both an outbreak study (November, 1998, to December, 2002) and a longitudinal study (1997 to 2018) in humans, as well as captures of Phlebotominae and potential reservoirs. Results: A total of 1,250 patients were registered; in the outbreaks, 190 (1998) to 249 cases (2002) were reported and inter-epidemic periods with 37 cases as an annual average; 68% of the patients were highland migrants who inhabited precarious housing near residual forests. The predominant sex was male (ratio 2:1); the most affected group (363/584 cases, 62%) was the economically active (15 to 49 years old); 124/584 cases (21%) were children under 15 years old, 33/584 of them were under 4 years old; 51 patients/584 (8.7%) had mucosal lesions. Leishmania (V.) braziliensis was isolated and characterized from mucous ulcers of sick dogs. Nyssomyia neivai, an abundant anthropophilic species incriminated as a probable vector, was captured. Conclusions: The initial 1997 leishmaniosis presence in the municipality of Bermejo had spread out over four municipalities in 2018 (Padcaya, Caraparí, Entre Ríos, and Yacuiba), northeast of the department of Tarija.

Introducción. En 1997, en el departamento de Tarija, Bolivia, situado en la frontera con Argentina, se notificó por primera vez la presencia de pacientes con úlceras en las partes descubiertas de la piel, cuyas características clínicas y epidemiológicas correspondían a leishmaniasis. Objetivo. Describir y comprobar la presencia de leishmaniasis en Tarija, sexto departamento endémico en Bolivia. Materiales y métodos. Se hizo un estudio del brote (noviembre de 1998 a diciembre de 2002) y un estudio longitudinal (1997 a 2018) en humanos; además, se capturaron Phlebotominae y potenciales reservorios. Resultados. Se registraron 1.250 pacientes de leishmaniasis; 190 y 249 casos, en los brotes de 1998 y 2002, respectivamente, con periodos interepidémicos de 37 casos como promedio anual. El 68 % de los enfermos eran pobladores migrantes del altiplano asentados en viviendas precarias cercanas al bosque residual; el sexo predominante fue el masculino (2/1). El grupo etario económicamente activo (15 a 49 años) fue el más afectado (363/584, 62 %). Hubo 124/584 (21 %) menores de 15 años, 33/584 de menos de cuatro años. En 51/584 (8,7 %) pacientes se presentaron lesiones mucosas. Se aisló y caracterizó Leishmania (V.) braziliensis de úlceras mucosas de perros enfermos y se capturó abundantemente la especie antropofílica Nyssomyia neivai, incriminada como probable vector. Conclusiones. En 1997 se comprobó por primera vez la presencia de leishmaniasis tegumentaria en el municipio de Bermejo y, en el 2018, ya se había extendido a cuatro municipios: Padcaya, Caraparí, Entre Ríos y Yacuiba, en dirección noreste del departamento de Tarija.

Variation of the Phlebotominae (Diptera: Psychodidae: Phlebotominae) Assemblage in Response to Land Use Changes in an Endemic Area of Leishmania Transmission in Northeast Argentina.

Leishmaniases are a global health problem and in Argentina are considered emerging diseases. The new transmission scenarios of tegumentary leishmaniasis are especially important given that large areas of forest are being transformed into rural and urban systems. The aim of this study was to analyze the impact of the construction of a large public building and a neighborhood on the assemblage of Phlebotominae in a rural area with forest remnants and to correlate the changes observed in the species assemblage with characteristics of the environment. Entomological surveys with light traps were conducted on the construction campus in the northeastern region of Argentina at six sites representing different environmental situations. Structural environmental characteristics and meteorological conditions were recorded and analyzed. At least 16 species of Phlebotominae sand flies were collected, the most prevalent being Nyssomyia whitmani (Antunes & Coutinho), followed by the genus Brumptomyia (França & Parrot) and Migonemyia migonei (França). Our study provides evidence of how the structure of the assemblages and prevalent species respond to anthropogenic disturbances. As the construction progressed, both Ny. whitmani and the genus Brumptomyia were favored. The genus Brumptomyia was favored at sites surrounded by high proportions of forest, within patches of remnant vegetation, and relatively far from anthropogenic disturbances, while Ny. whitmani, the main vector of tegumentary leishmaniasis in the region, increases their abundant at short and intermediate distances from vegetation margins and areas close to anthropogenic disturbances, therefore increasing the risk of human exposure to vectors.

Leishmaniasis en Bolivia, revisión y estado actual en Tarija, frontera con Argentina / Leishmaniasis en Bolivia. Comprehensive review and current status in Tarija, in the border with Argentina

Introducción. En 1997, en el departamento de Tarija, Bolivia, situado en la frontera con Argentina, se notificó por primera vez la presencia de pacientes con úlceras en las partes descubiertas de la piel, cuyas características clínicas y epidemiológicas correspondían a leishmaniasis. Objetivo. Describir y comprobar la presencia de leishmaniasis en Tarija, sexto departamento endémico en Bolivia. Materiales y métodos. Se hizo un estudio del brote (noviembre de 1998 a diciembre de 2002) y un estudio longitudinal (1997 a 2018) en humanos; además, se capturaron Phlebotominae y potenciales reservorios. Resultados. Se registraron 1.250 pacientes de leishmaniasis; 190 y 249 casos, en los brotes de 1998 y 2002, respectivamente, con periodos interepidémicos de 37 casos como promedio anual. El 68 % de los enfermos eran pobladores migrantes del altiplano asentados en viviendas precarias cercanas al bosque residual; el sexo predominante fue el masculino (2/1). El grupo etario económicamente activo (15 a 49 años) fue el más afectado (363/584, 62 %). Hubo 124/584 (21 %) menores de 15 años, 33/584 de menos de cuatro años. En 51/584 (8,7 %) pacientes se presentaron lesiones mucosas. Se aisló y caracterizó Leishmania (V.) braziliensis de úlceras mucosas de perros enfermos y se capturó abundantemente la especie antropofílica Nyssomyia neivai, incriminada como probable vector. Conclusiones. En 1997 se comprobó por primera vez la presencia de leishmaniasis tegumentaria en el municipio de Bermejo y, en el 2018, ya se había extendido a cuatro municipios: Padcaya, Caraparí, Entre Ríos y Yacuiba, en dirección noreste del departamento de Tarija.

Introduction: In the department of Tarija in the Bolivian-Argentine border, human cases with ulcers on uncovered parts of the skin plus clinical and epidemiological characteristics related to leishmaniasis were reported for the first time in 1997. Objective: To describe and to verify the presence of leishmaniasis in Tarija, sixth endemic department in Bolivia. Materials and methods: We conducted both an outbreak study (November, 1998, to December, 2002) and a longitudinal study (1997 to 2018) in humans, as well as captures of Phlebotominae and potential reservoirs. Results: A total of 1,250 patients were registered; in the outbreaks, 190 (1998) to 249 cases (2002) were reported and inter-epidemic periods with 37 cases as an annual average; 68% of the patients were highland migrants who inhabited precarious housing near residual forests. The predominant sex was male (ratio 2:1); the most affected group (363/584 cases, 62%) was the economically active (15 to 49 years old); 124/584 cases (21%) were children under 15 years old, 33/584 of them were under 4 years old; 51 patients/584 (8.7%) had mucosal lesions. Leishmania (V.) braziliensis was isolated and characterized from mucous ulcers of sick dogs. Nyssomyia neivai, an abundant anthropophilic species incriminated as a probable vector, was captured. Conclusions: The initial 1997 leishmaniosis presence in the municipality of Bermejo had spread out over four municipalities in 2018 (Padcaya, Caraparí, Entre Ríos, and Yacuiba), northeast of the department of Tarija.

Substantial reduction in child stunting is differentially associated to geographical and socioeconomic disparities in Misiones Province, Argentina.

OBJECTIVE: To estimate trends in the prevalence of child stunting in the population of children under 5 years of age covered by public health programmes, between 2009 and 2014 in Misiones, Argentina. METHODS: Using Bayesian model-based geostatistics, we evaluated 724 872 anthropometric measurements corresponding to 110 633 children. In order to identify disparities at local scale, we evaluated the hypotheses of a differential reduction of stunting according to the geographical location (at two-level spatial resolution) and to the socioeconomic level in a rural or urban environment. RESULTS: The prevalence of stunting had fallen significantly in the province overall. Sex and age defined gender disparities at individual level, and there were regional disparities with higher prevalence values in the north and northeast regions. In these areas, stunting decreased to a greater degree during the studied period, although the spatial pattern remained smoother. Stunting increased in peripheral urban and dispersed rural areas that are socioeconomically vulnerable. CONCLUSIONS: The spatial multi-level geostatistical estimates of child undernutrition provide a precision public health tool to target public policies to those populations with the greatest need, in order to reduce health disparities.

OBJECTIF: Estimer les tendances dans la prévalence du retard de croissance dans la population des enfants de moins de 5 ans couverts par les programmes de santé publique, entre 2009 et 2014 à Misiones, en Argentine. MÉTHODES: En utilisant la géostatistique basée sur un modèle bayésien, nous avons évalué 724.872 mesures anthropométriques correspondant à 110.633 enfants. Afin d'identifier les disparités à l'échelle locale, nous avons évalué les hypothèses d'une réduction différentielle du retard de croissance en fonction de la situation géographique (à une résolution spatiale à deux niveaux) et du niveau socioéconomique en milieu rural ou urbain. RÉSULTATS: La prévalence du retard de croissance avait considérablement diminué dans l'ensemble de la province. Le sexe et l'âge définissaient des disparités entre les sexes au niveau individuel, et il y avait des disparités régionales avec des valeurs de prévalence plus élevées dans les régions du nord et du nord-est. Dans ces régions, le retard de croissance a diminué plus fortement au cours de la période étudiée, bien que le modèle spatial soit resté plus lisse. Le retard de croissance a augmenté dans les zones urbaines périphériques et les zones rurales dispersées qui sont socioéconomiquement vulnérables. CONCLUSIONS: Les estimations géostatistiques spatiales à plusieurs niveaux de la sous-nutrition infantile fournissent un outil de santé publique de précision pour cibler les politiques publiques sur les populations qui en ont le plus besoin, afin de réduire les disparités en matière de santé.

Performance of rapid tests for canine visceral leishmaniasis diagnosis in Argentina.

To diagnose dogs infected by Leishmania infantum rK39 rapid diagnosis test is widely used in the Americas, while dual path platform (DPP) was recently adopted by Brazil. In this study we assessed the performance of rK39-RDT and DPP tests in recent urban transmission scenarios of Argentina. The sensitivity and specificity were evaluated with a sera panel and field samples, taken as true infected those from parasitological and/or PCR positive tests. Since none of these tests can be taken as a gold standard, the performance was also evaluated using Latent Class Analysis, a statistical modeling technique which allows to estimating sensitivity and specificity defining a latent class variable as the reference standard. The sensitivity of both tests in the panel was around 92% (symptomatic dogs 96%, asymptomatic 83%), while the sensitivity in field samples of rK39-RDT was 77%, and DPP 98% (mean in symptomatic dogs 89%, asymptomatic 82%). The specificity was similar for both tests and samples, around 98%. Therefore, these tests are acceptable for program dog population-based studies, as spatial stratification, focus intervention and follow up, and they could be used for individual screening and confirmation of clinical presumptive diagnosis in polysymptomatic dogs. The inability to discriminate between immunity and actual infectiousness suggest that a combination with other non-immunological based tests will be required for highly sensitive/specific diagnosis in order to targeting control measures in individual reservoirs from public health perspective, as for individual management from animal health perspective.

Para diagnosticar perros infectados por Leishmania infantum, en las Américas se utiliza ampliamente la prueba rápida rK39, mientras que DPP fue adoptado recientemente por Brasil. En este estudio se evaluó el desempeño de las pruebas rK39-RDT y DPP en escenarios de transmisión urbana reciente en Argentina. La sensibilidad y especificidad se evaluaron con un panel de sueros y muestras de campo, considerando muestras infectadas verdaderas aquellas con pruebas parasitológicas y/o de PCR positivas. Como ninguna de estas pruebas puede considerarse estándar de oro, el desempeño también se evaluó mediante análisis de clases latentes, una técnica de modelado estadístico que permite estimar sensibilidad y especificidad definiendo una variable de clase latente como estándar. La sensibilidad de ambas pruebas en el panel fue de alrededor del 92% (perros sintomáticos 96%, asintomáticos 83%), mientras que la sensibilidad en muestras de campo fue rK39-RDT: 77%, y DPP 98% (media en perros sintomáticos 89%, asintomáticos 82%). La especificidad fue similar para ambas pruebas y muestras, cerca de 98%. Por lo tanto, estas pruebas son aceptables para estudios programáticos caninos de base-poblacional, como estratificación espacial, intervención de foco y seguimiento, y podrían utilizarse para el tamizaje individual y la confirmación del diagnóstico clínico presuntivo en perros poli-sintomáticos. La incapacidad de discriminar entre inmunidad e infectividad real sugiere que se requerirá una combinación con otras pruebas, de base no inmunológica, para un diagnóstico suficientemente sensible/específico que permita definir las medidas de control en reservorios individuales, tanto para salud pública, como para la gestión individual en salud animal.

Performance of rapid tests for canine visceral leishmaniasis diagnosis in Argentina

To diagnose dogs infected by Leishmania infantum rK39 rapid diagnosis test is widely used in the Americas, while dual path platform (DPP) was recently adopted by Brazil. In this study we assessed the performance of rK39-RDT and DPP tests in recent urban transmission scenarios of Argentina. The sensitivity and specificity were evaluated with a sera panel and field samples, taken as true infected those from parasitological and/or PCR positive tests. Since none of these tests can be taken as a gold standard, the performance was also evaluated using Latent Class Analysis, a statistical modeling technique which allows to estimating sensitivity and specificity defining a latent class variable as the reference standard. The sensitivity of both tests in the panel was around 92% (symptomatic dogs 96%, asymptomatic 83%), while the sensitivity in field samples of rK39-RDT was 77%, and DPP 98% (mean in symptomatic dogs 89%, asymptomatic 82%). The specificity was similar for both tests and samples, around 98%. Therefore, these tests are acceptable for program dog population-based studies, as spatial stratification, focus intervention and follow up, and they could be used for individual screening and confirmation of clinical presumptive diagnosis in polysymptomatic dogs. The inability to discriminate between immunity and actual infectiousness suggest that a combination with other non-immunological based tests will be required for highly sensitive/specific diagnosis in order to targeting control measures in individual reservoirs from public health perspective, as for individual management from animal health perspective.

Para diagnosticar perros infectados por Leishmania infantum, en las Américas se utiliza ampliamente la prueba rápida rK39, mientras que DPP fue adoptado recientemente por Brasil. En este estudio se evaluó el desempeño de las pruebas rK39-RDT y DPP en escenarios de transmisión urbana reciente en Argentina. La sensibilidad y especificidad se evaluaron con un panel de sueros y muestras de campo, considerando muestras infectadas verdaderas aquellas con pruebas parasitológicas y/o de PCR positivas. Como ninguna de estas pruebas puede considerarse estándar de oro, el desempeño también se evaluó mediante análisis de clases latentes, una técnica de modelado estadístico que permite estimar sensibilidad y especificidad definiendo una variable de clase latente como estándar. La sensibilidad de ambas pruebas en el panel fue de alrededor del 92% (perros sintomáticos 96%, asintomáticos 83%), mientras que la sensibilidad en muestras de campo fue rK39-RDT: 77%, y DPP 98% (media en perros sintomáticos 89%, asintomáticos 82%). La especificidad fue similar para ambas pruebas y muestras, cerca de 98%. Por lo tanto, estas pruebas son aceptables para estudios programáticos caninos de base-poblacional, como estratificación espacial, intervención de foco y seguimiento, y podrían utilizarse para el tamizaje individual y la confirmación del diagnóstico clínico presuntivo en perros poli-sintomáticos. La incapacidad de discriminar entre inmunidad e infectividad real sugiere que se requerirá una combinación con otras pruebas, de base no inmunológica, para un diagnóstico suficientemente sensible/específico que permita definir las medidas de control en reservorios individuales, tanto para salud pública, como para la gestión individual en salud animal.

Molecular detection and identification of Bartonella in the cat flea Ctenocephalides felis felis collected from companion animals in a border area in northeastern Argentina.

Molecular methods were used to detect and identify Bartonella species in the cat fleas Ctenocephalides felis felis from Puerto Iguazú, a border area in northeastern Argentina. The fleas were collected from 12 household animals, 9 dogs (Canis lupus familiaris) and 3 cats (Felis silvestris catus) during July 2016. Out of 15C. f. felis analyzed for PCR, only one flea collected from a cat was positive (6.66%) in screened for Bartonella spp. based on the gltA gene. Bartonella clarridgeiae was identified in the genetic analyses, this specimen clustered monophyletically with others B. clarridgeiae isolated from different geographical origins (1.0 PP), even, all shared the same haplotype. The results obtained provide evidence of the presence of B. clarridgeiae in cat fleas from Argentina suggesting the probable presence of related flea-borne diseases in the region and the role of cat fleas in the transmission of Bartonella among mammals including humans.

Visceral Leishmaniasis in Bolivia: Current Status

Abstract INTRODUCTION In Bolivia, before 1982 there were no records of visceral leishmaniasis (VL) cases that would allow us to review and describe the temporospatial occurrence of VL by ecoregions in provinces and departments of Bolivia to evaluate its impact on public health, risk of outbreaks, or dispersion. METHODS This update on VL in Bolivia is based on research, reviews, and retrospective literature analyses of online data and libraries and institutional reports, from 1939 to the present. RESULTS In Bolivia, 56 cases of VL have been reported. Until 2014, only three endemic departments had been identified (La Paz, Santa Cruz, and Tarija). Since then, further cases have been recorded in Pando, Cochabamba, and Beni, and in Chuquisaca in 2015. In Yungas, a VL focus was confirmed by isolating and comparing parasites from human and dog cases, and from the Lu. longipalpis vector. VL cases from seven departments, involving 12 different ecoregions were located within the Amazon and Plata basins. CONCLUSIONS We confirmed that dogs are its primary reservoir, and Lutzomyia longipalpis is its main vector (currently dispersed in six departments). The primary vectors in areas where Lutzomyia longipalpis is absent are Migonemyia migonei and Lutzomyia cruzi.

Optimization of DNA Extraction from Individual Sand Flies for PCR Amplification.

Numerous protocols have been published for extracting DNA from phlebotomines. Nevertheless, their small size is generally an issue in terms of yield, efficiency, and purity, for large-scale individual sand fly DNA extractions when using traditional methods. Even though this can be circumvented with commercial kits, these are generally cost-prohibitive for developing countries. We encountered these limitations when analyzing field-collected Lutzomyia spp. by polymerase chain reaction (PCR) and, for this reason, we evaluated various modifications on a previously published protocol, the most significant of which was a different lysis buffer that contained Ca2+ (buffer TESCa). This ion protects proteinase K against autolysis, increases its thermal stability, and could have a regulatory function for its substrate-binding site. Individual sand fly DNA extraction success was confirmed by amplification reactions using internal control primers that amplify a fragment of the cacophony gene. To the best of our knowledge, this is the first time a lysis buffer containing Ca2+ has been reported for the extraction of DNA from sand flies.

Genetic diversity, phylogeography and molecular clock of the Lutzomyia longipalpis complex (Diptera: Psychodidae).

BACKGROUND: The Lutzomyia longipalpis complex has a wide but discontinuous distribution in Latin America, extending throughout the Neotropical realm between Mexico and northern Argentina and Uruguay. In the Americas, this sandfly is the main vector of Leishmania infantum, the parasite responsible for Visceral Leishmaniasis (VL). The Lu. longipalpis complex consists of at least four sibling species, however, there is no current consensus on the number of haplogroups, or on their divergence. Particularly in Argentina, there have been few genetic analyses of Lu. longipalpis, despite its southern expansion and recent colonization of urban environments. The aim of this study was to analyze the genetic diversity and structure of Lu. longipalpis from Argentina, and to integrate these data to re-evaluate the phylogeography of the Lu. longipalpis complex using mitochondrial markers at a Latin American scale. METHODOLOGY/PRINCIPAL FINDINGS: Genetic diversity was estimated from six sites in Argentina, using a fragment of the ND4 and the 3´ extreme of the cyt b genes. Greatest genetic diversity was found in Tartagal, Santo Tomé and San Ignacio. There was high genetic differentiation of Lu. longipalpis in Argentina using both markers: ND4 (FST = 0.452, p < 0.0001), cyt b (FST = 0.201, p < 0.0001). Genetic and spatial Geneland analyses reveal the existence of two primary genetic clusters in Argentina, cluster 1: Tartagal, Santo Tomé, and San Ignacio; cluster 2: Puerto Iguazú, Clorinda, and Corrientes city. Phylogeographic analyses using ND4 and cyt b gene sequences available in GenBank from diverse geographic sites suggest greater divergence than previously reported. At least eight haplogroups (three of these identified in Argentina), each separated by multiple mutational steps using the ND4, are differentiated across the Neotropical realm. The divergence of the Lu. longipalpis complex from its most recent common ancestor (MRCA) was estimated to have occurred 0.70 MYA (95% HPD interval = 0.48-0.99 MYA). CONCLUSIONS/SIGNIFICANCE: This study provides new evidence supporting two Lu. longipalpis genetic clusters and three of the total eight haplogroups circulating in Argentina. There was a high level of phylogeographic divergence among the eight haplogroups of the Lu. longipalpis complex across the Neotropical realm. These findings suggest the need to analyze vector competence, among other parameters intrinsic to a zoonosis, according to vector haplogroup, and to consider these in the design and surveillance of vector and transmission control strategies.

Considerations on the Species Complex of the Cortelezzii series (Diptera: Psychodidae) and Description of Evandromyia chacuensis sp. nov., a New Phlebotomine Species of the Chaco Region, Argentina.

A new species of phlebotomine sand fly is described and illustrated based on male and female specimens collected in rural and semi-urban areas of the Chaco Province, Argentina. A separation of the Cortelezzii series in two species complexes is proposed, as is a species key for its identification. The morphological characters of the new species permit its inclusion in the Cortelezzii series of genus Evandromyia, subgenus Barrettomyia (Diptera: Psychodidae). The species is named Evandromyia chacuensis sp. nov. Szelag, Rosa, Galati, Andrade Fhilo & Salomón (Diptera: Psychodidae).

Environmental and socio-demographic individual, family and neighborhood factors associated with children intestinal parasitoses at Iguazú, in the subtropical northern border of Argentina.

BACKGROUND: Intestinal parasitoses are a major concern for public health, especially in children from middle and low-income populations of tropical and subtropical areas. We examined the presence and co-infection of parasites in humans as well as parasitic environmental contamination in Puerto Iguazú, Argentina. We explored the environmental and socio-demographic characteristics of the persistence of parasites in children and their environment. METHODOLOGY/PRINCIPAL FINDINGS: This cross-section survey was conducted among children population comprised into the area of the public health care centers of Iguazú during June 2013 to May 2016. Copro-parasitological status of 483 asymptomatic children was assessed. Simultaneously, a design-based sampling of 744 soil samples and 530 dog feces was used for characterize the environmental contamination. The 71.5% of these sites were contaminated. Sixteen genera were detected in the environment being hookworms (62.0%) the main pathogens group detected followed by Toxocara spp (16.3%), Trichuris spp (15.2%) and Giardia (6.5%). Total children prevalence raised 58.8%, detecting twelve genera of parasite with Giardia intestinalis as the most prevalent pathogen (29.0%) followed by Enterobius vermicularis (23.0%), Hymenolepis nana (12.4%) and hookworms (4.4%). Through questionnaires and census data, we characterized the socio-demographics conditions at an individual, family and neighborhood levels. A multi-level analysis including environmental contamination data showed that the ´presence of parasites´ was mostly determined by individual (e.g. age, playing habits, previous treatment) and household level (e.g. UBN, WASH, mother's literacy) determinants. Remarkably, to define the level of 'parasite co-infection', besides individual and household characteristics, environmental factors at a neighborhood level were important. CONCLUSION/SIGNIFICANCE: Our work represents the major survey of intestinal parasites in human and environmental samples developed in the region. High prevalence was detected in the children population as well as in their environment. This work shows the importance of considering and promoting multi-level actions over the identified determinants to face this public health problem from integrative approaches.

Zika Virus: What Have We Learnt Since the Start of the Recent Epidemic?

Zika is a viral disease transmitted mainly by mosquitoes of the genus Aedes. In recent years, it has expanded geographically, changing from an endemic mosquito-borne disease across equatorial Asia and Africa, to an epidemic disease causing large outbreaks in several areas of the world. With the recent Zika virus (ZIKV) outbreaks in the Americas, the disease has become a focus of attention of public health agencies and of the international research community, especially due to an association with neurological disorders in adults and to the severe neurological and ophthalmological abnormalities found in fetuses and newborns of mothers exposed to ZIKV during pregnancy. A large number of studies have been published in the last 3 years, revealing the structure of the virus, how it is transmitted and how it affects human cells. Many different animal models have been developed, which recapitulate several features of ZIKV disease and its neurological consequences. Moreover, several vaccine candidates are now in active preclinical development, and three of them have already entered phase I clinical trials. Likewise, many different compounds targeting viral and cellular components are being tested in in vitro and in experimental animal models. This review aims to discuss the current state of this rapidly growing literature from a multidisciplinary perspective, as well as to present an overview of the public health response to Zika and of the perspectives for the prevention and treatment of this disease.

Spatial population dynamics and temporal analysis of the distribution of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) in the city of Clorinda, Formosa, Argentina.

BACKGROUND: Lutzomyia longipalpis, the vector for the causal agent of visceral leishmaniasis (VL), has extended its distribution in the southern cone in the Americas. The first urban record of Lu. longipalpis in Argentina was from the City of Clorinda in 2004. The aim of this study was to analyse the monthly distribution and abundance of Lu. longipalpis and to evaluate its association with environmental and climatic variables in Clorinda City, Province of Formosa. METHODS: Phlebotominae sampling was performed using CDC light mini-traps that were placed in different sites of the city between January 2012 and December 2013. Environmental variables including the normalised difference vegetation index, normalized difference water index, land surface temperature and precipitation were evaluated using a spatiotemporal model. RESULTS: A total of 4996 phlebotomine sandflies were captured during the study period, and eight species were reported: Lu. longipalpis, Migonemyia migonei, Nyssomyia whitmani, Ny. neivai, Brumptomyia guimaraesi, Evandromyia cortelezzii/sallesi, Psathyromyia bigeniculata and Expapillata firmatoi. This is the first urban record of Ex. firmatoi in Argentina. Lutzomyia longipalpis was the most abundant species between 2012 and 2013, and it appeared in all the sampled sites. Moreover, the model applied showed that ground humidity and temperature were significantly associated with the abundance of Lu. longipalpis. CONCLUSIONS: This longitudinal approach at city scale allows for modelling that explains more than 60% of the temporal variability of the abundance of Lu. longipalpis based exclusively on satellite obtained data. The results support the hypothesis of steady 'hot spots' of abundance with time, while other sites could change its abundance due to eventual microenvironment changes. The Lu. longipalpis abundance driving factors are breeding site-related variables, highlighting the importance both for modelling and surveillance to use lag data.