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.

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.

[Dengue vaccines. A reality for Argentina?]. / Las vacunas contra el dengue ¿Una realidad para la Argentina?

Dengue outbreaks have occurred yearly in Argentina since 1998. A number of candidate vaccines have been tested in endemic countries. The most advanced one was licensed in three countries of Latin America for children over 9 years of age. In the present article the benefits and drawbacks of these vaccines as well as the challenges for the implementation of a vaccination strategy in Argentina are discussed. Furthermore, a risk stratification strategy with new criteria and a multidisciplinary vision is suggested as a possible path for the assessment of the pertinence of a vaccination program in areas showing the highest risk of dengue transmission and/or for people at the greatest risk of developing severe dengue. It is also suggested that the definition regarding the status of endemicity should take into account the local realities. Finally, this paper proposes a broad discussion on the evidences, the expected impact and instrumental aspects that would be involved in the incorporation of a dengue vaccine, marketed or in development, into the national immunization program, and especially which subpopulation should be targeted for the immunization strategy to be cost-effective.

Diphyllobothrium sp. in Canis familiaris from the subtropical area of Argentina (Puerto Iguazú, Misiones).

This paper reports the first finding of Diphyllobothrium sp. eggs in Canis familiaris (domestic dog) from Puerto Iguazú, a subtropical city of Misiones province, Argentina. In 2013, two positive cases of Diphyllobothrium sp. eggs were detected during an annual parasitological survey of dogs. Dog feces were collected in vials containing 10% formalin and processed using Telemann's sedimentation and Sheather's flotation techniques. The two cases were detected in rural areas of the municipality. Since Misiones is not a part of the endemic area of diphyllobothriasis and given the fact that it is located in the three-border area of Argentina, Brazil and Paraguay, we consider this finding of great importance to public health. We stress the need for updating the current knowledge about the life cycle of these parasites considering the range of intermediate and definitive hosts, their zoonotic potential, and the epidemiological situation in non-endemic areas.

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.

[Visceral leishmaniasis in Argentina. Cases notification and distribution of vectors (2006-2012)]. / Leishmaniasis visceral en la Argentina. Notificación y situación vectorial (2006-2012).

Visceral leishmaniasis (VL) is a notifiable disease in Argentina. Every new case diagnosed must be reported to the National Health Surveillance System. In the period 2006-2012, a total of 103 cases have been notified in four provinces: Entre Rios, Santa Fe, Misiones and Santiago del Estero. Misiones has the largest amount of cases: a total of 80, detected in 15 towns from May 2006 to July 2012. The highest incidence is observed in children (less than 15 years of age). In 2004 the vector Lutzomyia longipalpis was detected for the first time in Formosa Province. Cases of human and canine VL, as well as the presence of the canine vector were reported in 2006 in the province of Misiones, and in 2008-2009 in the province of Corrientes. In Santiago del Estero cases of human and canine VL (2008) were possibly associated with secondary vectors. In 2010 Lu longipalpis was found in Entre Rios, showing its dispersal to the south. Between January and April 2011 a systematic search of the VL vector was carried out in 13 towns, in Entre Ríos, Santa Fe and Santiago del Estero Provinces (275 traps/night). The colonization of the vector Lu longipalpis was proved in Chajarí town (Entre Rios), but not in Santiago del Estero Province, where Migonemyia migonei appears as a putative LV vector.

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.

[Distribution of Lutzomyia longipalpis in the Chaco region, Argentina, 2010]. / Distribución de Lutzomyia longipalpis en el Chaco Argentino, 2010.

The appearance of the vector of visceral leishmaniasis (VL) Lutzomyia longipalpis in the province of Formosa in 2004 was associated with urban epidemic risk for the first time in Argentina. During 2006, vectors, canine and human cases of VL were reported in the province of Misiones, and in summer 2008-2009, in the province of Corrientes. In Santiago del Estero province in 2008, cases of human and canine VL were associated with secondary vectors. Therefore, with the aim to know the current distribution of VL risk in the Chaco region, systematic captures of the vector were performed between January and April 2010 in 30 localities of Formosa and Chaco, and in the city of Reconquista, province of Santa Fe (224 traps/night). Lu. longipalpis was reported for the first time in the cities of Resistencia and Puerto Antequera (Chaco). Clorinda and Puerto Pilcomayo (Formosa) are the localities where the traps with more individuals were obtained, 158 and 241 Lu. longipalpis trap / site / night respectively. These results showed that the vector of urban epidemic VL is still spreading in Argentina, and already reached the province of Chaco. Sporadic cases reported in the Chaco region, transmitted by secondary vectors as Lu. migonei would also increase due to intensified surveillance, and the dispersion of the parasite associated with the transit of asymptomatic or symptomatic infected dogs.

[Distribution of Lutzomyia longipalpis in the Argentine Mesopotamia, 2010]. / Distribución de Lutzomyia Longipalpis en la Mesopotamia Argentina, 2010.

The first case of visceral leishmaniasis (VL) in Argentina was reported in 2006 in Posadas, Misiones. During the summer 2008-2009 Lutzomyia longipalpis, the VL vector, and canine VL cases were already spread along the province of Corrientes. In order to know the distribution of VL risk, systematic captures of the vector were performed between February and March 2010, in 18 areas of the provinces of Entre Ríos and Corrientes, and the city of Puerto Iguazú, Misiones, with a total of 313 traps/night. We confirmed the presence of Lu. longipalpis, for the first time in Chajarí (Entre Ríos), Alvear, La Cruz, Curuzú Cuatiá and Bella Vista (Corrientes), and Puerto Iguazú (Misiones). In Santo Tome and Monte Caseros (Corrientes), where the vector had been previously reported, traps with more samples were obtained with 830 and 126 Lu. Longipalpis trap/site/night respectively. These results show that the vector of urban VL continues spreading in the Argentine territory. Simultaneously, the spread of the parasite and the resulting human VL cases are associated with the dispersion of reservoirs, infected dogs, with or without clinical symptoms or signs, due to human transit.

Sex pheromone and period gene characterization of Lutzomyia longipalpis sensu lato (Lutz & Neiva) (Diptera: Psychodidae) from Posadas, Argentina.

Lutzomyia longipalpis s.l. is the primary vector of Leishmania (L.) infantum in the New World. In this study, male Lutzomyia longipalpis specimens from Posadas, Argentina were characterized for two polymorphic markers: the male sex pheromone and the period (per) gene. The male sex pheromone was identified as (S)-9-methylgermacrene-B, the same compound produced by Lu. longipalpis from Paraguay and many populations from Brazil. The analysis of per gene sequences revealed that the population from Argentina is significantly differentiated from previously studied Brazilian populations. Marker studies could contribute to the understanding of the distribution and spread of urban American visceral leishmaniasis, thus aiding in the design of regional surveillance and control strategies.

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.

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.

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.

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.

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.

A new venomous scorpion responsible for severe envenomation in Argentina: Tityus confluens.

In Argentina the scorpions of medical importance belong to the genus Tityus (T.), particularly the species T. trivittatus, the only scorpion whose sting is recognized to be associated with severe human envenoming and death. This genus is distributed from the north of the Patagonian region to the center and some provinces in the north of the country. During the period 2003-2006 four children died following scorpion stings, of which one was certainly and three were probably by T. confluens. In 2006, in the province of Tucumán, a girl died by scorpion envenoming and the scorpion responsible for the death, found in her shoe, was T. confluens. We thus studied the toxicity of venom gland homogenates from T. confluens from the provinces of Jujuy and Catamarca, and of crude venom from specimens from Catamarca and the province of La Rioja. The lethal potencies of the telson homogenates were 7.0 and 18.6microg/g for Jujuy and Catamarca, respectively, while the lethal potency of the crude venom was 0.7microg/g. Injected mice showed generalized congestion and hepatic lesions. Pancreatic damage was observed in some animals. Lungs showed congestion and foci of hemorrhage and mild edema. The heart showed injury in the muscular fibers. The venom showed high reactivity against anti-T. trivittatus antivenom and against two anti-T. serrulatus antivenoms. The anti-T. trivittatus antivenom neutralized the lethal activity of T. confluens venom. In addition, the venom reacted very slightly against an anti-Centruroides antivenom. Therefore, the stings of this scorpion must be considered of risk for humans to the same degree as the stings of T. trivittatus.

Visceral leishmaniasis in border areas: clustered distribution of phlebotomine sand flies in Clorinda, Argentina.

Three years after the first report of Lutzomyia longipalpis in Clorinda, Argentina, a border city near Asunción, Paraguay, the city was surveyed again. Lu. longipalpis was found clustered in the same neighbourhoods in 2007 as in 2004, even though the scattered distribution of canine visceral leishmaniasis was more related to the traffic of dogs through the border.

[Distribution of vectors of visceral leishmaniasis in the Province of Corrientes, 2008]. / Distribución de vectores de leishmaniasis visceral en la Provincia de Corrientes, 2008.

Visceral leishmaniasis (VL) is a relevant parasitic disease in public health, produced by Leishmania infantum chagasi. Since the urbanization and emergence in Southern Brazil and Paraguay, the vector Lutzomyia longipalpis in Formosa, 2004, and the first human visceral leishmaniasis case in Misiones, 2006, have been reported in Argentina. Due to the reports of canine VL, a search of the vector in the Province of Corrientes, contiguous to Misiones, was performed during December 2008. Standarized trapping detected 376 Lu. longipalpis in Ituzaingó, Virasoro, Santo Tomé, Garruchos, Riachuelo, Corrientes and Monte Caseros localities. The risk of autochtonous vectorial transmission was then confirmed in the Province of Corrientes. The distribution of vectors in populated urban areas, with intense transit of canine reservoirs from localities with high transmission, and the existence of infected reservoirs, also implies epidemic risk.

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.