One piece of the puzzle: Modeling vector presence and environment reveals seasonality, distribution, and prevalence of sandflies and Leishmania in an expansion area.

The recent geographic spread of Leishmania infantum along the borders of Argentina, Brazil and Paraguay has been highlighted. In our previous study, Lutzomyia longipalpis was found in 55 of 123 patches surveyed, and in some patches, sandflies were found at higher densities, forming hotspots. Based on the One Health approach, we investigated the seasonality of the vector, the presence of parasite DNA, and the environmental factors that contribute to vector and parasite dispersal in these previously described hotspots in Foz do Iguaçu, Brazil. Entomological surveys were conducted monthly for one year. Fourteen hotspots peridomicile and six intradomicile were sampled. PCR was used to assess the prevalence of Leishmania DNA in sandflies. Zero-inflated negative binomial regression was used to determine the association of micro- and mesoscale environmental variables with the occurrence and abundance of the three most abundant sandfly species sampled. A total of 3543 species were captured, with Lutzomyia longipalpis being the predominant species (71.78%) of the 13 species found. Evandromyia edwardsi, Expapillata firmatoi, Micropygomyia ferreirana and Pintomyia christenseni were reported for the first time in the region. NDVI, distance to water, precipitation, west-to-east wind, wind speed, maximum and minimum relative humidity, and sex were significant variables associated with vector presence/abundance in the environment. Vector presence/abundance in the peridomicile was associated with precipitation, altitude, maximum temperature, minimum and maximum relative humidity, west-to-east wind, wind speed, and sex. Leishmania DNA was detected in an average of 21% of Lu. longipalpis throughout the year. Vector abundance is concentrated in urban and peri-urban areas, with some specimens present in different parts of the city and some sites with high vector abundance. This distribution suggests that the risk of actual contact between humans and parasite vectors in urban areas during the epidemic period is associated with patches of peri-urban vegetation and then extends into urban areas.

GENETIC DIVERSITY OF Lutzomyia (Nyssomyia) intermedia IN AN ENDEMIC AREA OF AMERICAN CUTANEOUS LEISHMANIASIS, STATE OF PARANÁ, BRAZIL / Diversidad genética de Lutzomyia (Nyssomyia) intermedia en un área endémica de leishmaniasis cutánea americana del estado de Paraná, Brasil

ABSTRACT Lutzomyia intermedia (Diptera: Psychodidae) features as one of the main vectors that are involved in the transmission of American cutaneous leishmaniasis (ACL) in the Neotropical region. However, genetic studies involving this taxon are still incipient and important for understanding the level of variability of different populations, their role, and implications as vectors. The aim of this study was to determine the level of genetic diversity of L. intermedia present in the Ribeira River Valley, an area of ACL transmission in the state of Paraná, Brazil, through the Random amplified polymorphic DNA (RAPD). Two municipalities were chosen to collect sand flies: Cerro Azul (new transmission area of the ACL) and Adrianópolis (endemic area of the ACL). The insects were captured in the house, in the peridomicile and in the wild (forest). Two of the used markers made it possible to estimate the polymorphism of the studied populations, resulting in 40 genotypes, most of them from peridomicile. The dendrogram generated by the analysis with the primer A10 showed different degrees of similarity, suggesting that there may be gene flow in the studied populations. The Principal Coordinate Analysis (PCO) with the A2 primer, was useful in grouping L. intermedia according to its ecological and geographical origin. There was no distinction between the lineages composing the L. intermedia complex. The results of this study, with the record of great genotypic diversity in L. intermedia, may contribute to explain the maintenance of the life cycle of Leishmania braziliensis (Kinetoplastida: Trypanosomatidae) in the region.

RESUMEN Lutzomyia intermedia (Diptera: Psychodidae) es uno de los principales vectores que participan en la transmisión de leishmaniasis cutánea americana (LCA) en la región Neotropical. A pesar de que aún los estudios genéticos que involucran a este taxón son incipientes, tienen una gran importancia para comprender el nivel de variabilidad de las diferentes poblaciones y sus implicaciones en su papel vectorial. El objetivo de este estudio fue determinar el nivel de diversidad genética de L. intermedia presente en el Valle del Río Ribeira, área de transmisión de LCA en el estado de Paraná, Brasil, mediante RAPD (ADN polimórfico amplificado aleatoriamente). Los flebótomos fueron recolectados en los municipios Cerro Azul (nueva área de transmisión de LCA) y Adrianópolis (área endémica de LCA), donde fueron capturados en ambientes residenciales, en el peridomicilio y en el bosque. Dos de los marcadores utilizados permitieron estimar el polimorfismo en las poblaciones estudiadas con la obtención de 40 genotipos, la mayoría de ellos en el peridomicilio. El dendrograma generado por el análisis con el cebador A10 mostró diferentes grados de similitud, lo que sugiere que puede haber flujo gènico en las poblaciones. El Análisis de Coordenadas Principales (PCO) con el cebador A2 fue útil para agrupar L. intermedia según su origen ecológico y geográfico. No hubo distinción entre los linajes que componen el complejo L. intermedia. Los resultados de este estudio, con el registro de gran diversidad genotipica en L. intermedia, pueden contribuir a explicar el mantenimiento del ciclo biológico de Leishmania braziliensis (Kinetoplastida: Trypanosomatidae) en la región.

Abundancia y diversidad de vectores (Diptera: Psychodidae) en área de transmisión antigua de Leishmaniasis Cutánea en el nuevo mundo después de la construcción del gasoducto Bolivia-Brasil / Abundance and diversity of vectors (Diptera: Psychodidae) in an old transmission area of cutaneous leishmaniasis in the new world after Bolivia-Brazil gas pipeline construction

The objective of this study was to determine the abundance and diversity of the species and their presumed vectorial role in the transmission of Cutaneous Leishmaniasis (CL). Research on sand flies (Diptera: Psychodidae) was carried out in two municipalities in the Ribeira River Valley in the State of Paraná, southern Brazil. One, Adrianópolis, is an endemic area of CL and the other is a recent outbreak area (Cerro Azul). A total of 432 specimens were collected from residential, peri-domestic and wild environments. According to the data obtained, for each ecotope studied, the statistical analysis has shown that two lines of the Lutzomyia intermedia population coexist and that the quantity of specimens from L. intermedia s.l. is significantly different from L. intermedia s.s. in all environments (χ2=9.943; DF=2; p=0.07). The prevalence of L. intermedia (93.28%) in the ecotopes studied suggests that it is the main vector of leishmaniasis. The succession of vector species and their involvement in the epidemiological cycle of L. braziliensis is discussed in this article(AU)

El objetivo de este trabajo fue determinar la abundancia y diversidad de las especies y su supuesto papel vectorial en la transmisión de Leishmaniasis Cutánea (LC). La investigación sobre flebótomos (Diptera: Psychodidae) se llevó a cabo en dos municipios en el Valle del Río Ribeira en el Estado de Paraná, sur de Brasil. Las regiones estudiadas fueron Adrianópolis, área endémica de LC, y Cerro Azul, que se reporta como zona de brote reciente. Se recolectaron un total de 432 especímenes de ambientes residenciales, peri-domésticos y silvestres. El análisis estadístico realizado, a partir de los datos obtenidos para cada ecotopo estudiado, mostró que coexistían dos linajes de la población de Lutzomyia intermedia y que la cantidad de especímenes de L. intermedia s.l. fue significativamente diferente de L. intermedia s.s. en todos los entornos (χ2=9,943; GL=2; p=0,07). La prevalencia de L. intermedia (93,28%) en los ecotopos estudiados sugiere que es el principal vector de la leishmaniasis. La sucesión de especies de vectores y su participación en el ciclo epidemiológico de L. braziliensis se discute en este artículo(AU)

Hidden danger: Unexpected scenario in the vector-parasite dynamics of leishmaniases in the Brazil side of triple border (Argentina, Brazil and Paraguay).

Every year about 3 million tourists from around the world visit Brazil, Argentina and Paraguay´s triple border region where the Iguaçu Falls are located. Unfortunately, in recent years an increasing number of autochthonous canine and human visceral leishmaniasis (VL) cases have been reported. The parasite is Leishmania (Leishmania) infantum and it is transmitted by sand flies (Phlebotominae). To assess the risk factors favorable for the establishment and spread of potential vectors the Centers for Disease Control and Prevention light trap (CDC-light trap) collections were made in the Foz do Iguaçu (FI) and Santa Terezinha de Itaipu (STI) townships and along two transects between them. Our study determined the Phlebotominae fauna, the factors that affect the presence and abundance of Lutzomyia longipalpis and Nyssomyia whitmani, the presence of L. infantum in different sand fly species and which Leishmania species are present in this region. Lutzomyia longipalpis was the prevalent species and its distribution was related to the abundance of dogs. Leishmania infantum was found in Lu. longipalpis, Ny. whitmani, Ny. neivai and a Lutzomyia sp. All the results are discussed within the Stockholm Paradigm and focus on their importance in the elaboration of public health policies in international border areas. This region has all the properties of stable VL endemic foci that can serve as a source of the disease for neighboring municipalities, states and countries. Most of the urban areas of tropical America are propitious for Lu. longipalpis establishment and have large dog populations. Pan American Health Organization´s initiative in supporting the public health policies in the border areas of this study is crucial and laudable. However, if stakeholders do not act quickly in controlling VL in this region, the scenario will inevitable become worse. Moreover, L. (Viannia) braziliensis found in this study supports the need to develop public health policies to avoid the spread of cutaneous leishmaniasis. The consequences of socioeconomic attributes, boundaries and frontiers on the spread of diseases cannot be neglected. For an efficient control, it is essential that urban planning is articulated with the neighboring cities.

Cutaneous leishmaniosis in naturally infected dogs in Paraná, Brazil, and the epidemiological implications of Leishmania (Viannia) braziliensis detection in internal organs and intact skin.

Environmental changes have occurred over the years, altering the eco-epidemiological pattern of leishmaniosis in the State of Paraná, Brazil, involving the pillars of the cycle (parasite, vectors, reservoir, and environment) and their interaction. Much has been discussed about the dog's role as a reservoir of the Leishmania (Viannia) braziliensis Vianna, 1911 transmission cycle. However, this question remains unanswered. The purpose of this study was to investigate, using parasitological and molecular methods, different samples in eight naturally infected dogs from an endemic rural locality where only L. (V.) braziliensis is present, and where human cases have been previously notified. Blood and biopsied organ samples from naturally infected dogs were analyzed by culture media, PCR, random amplified polymorphic DNA and sequencing methodologies. Only skin lesions from all dogs yielded positive cultures and when PCR was performed, L. (V.) braziliensis DNA was amplified from intact skin, peripheral blood, bone marrow, spleen, liver and lymph nodes. RAPD was also applied to isolates from the skin lesions, exhibiting the genetic variability of the parasite identified. To confirm which species of Leishmania was amplified in PCR, the sequencing method was performed, verifying 100% similarity with the Viannia subgenus. This study showed that L. (V.) braziliensis can spread to other sites besides the ulcerous lesions, such as intact skin, peripheral blood and internal organs, making it possibility for dogs to serve as active sources of parasite transmission. For definitive proof, xenodiagnostic test on intact skin of infected dogs, should be done.

A mesmice identitária: a (im)possibilidade de emancipação na política de assistência social / La mismidad de identidad: la (im)posibilidad de emancipación en la política de asistencia social / The sameness of identity: the (im)possibility of emancipation in social assistance policy

Resumo O artigo reflete resultados de duas pesquisas de iniciação científica que tiveram o escopo basilar da teoria psicossocial proposta pelo professor Ciampa, que concebe a identidade enquanto metamorfose humana em busca de emancipação. Esse enfoque teórico metodológico possibilitou um entendimento conceitual crítico sobre aspectos relevantes da política nacional de assistência social e a inserção da psicologia nessa produção. Para o intento foi pesquisada a construção identitária de psicólogos e cidadãos usuários dos Centros de Referência de Assistência Social (CRAS), utilizando-se métodos qualitativos com ênfase nas entrevistas de história de vida. Dentre os fragmentos analisados destaca-se a percepção do psicólogo sobre sua identidade profissional (angústias e dificuldades sistêmicas que entravam suas práticas emancipatórias); em relação aos cidadãos usuários, percebe-se a falta de compreensão sobre o ideário da política, o não reconhecimento da psicologia enquanto prática comunitária e o conformismo e a resignação diante das precariedades vividas.

Resumen Este artículo analiza los resultados de dos investigaciones que se apoyaron en la teoría psicosocial propuesta por Ciampa, que concibe la identidad como metamorfosis humana en la búsqueda de emancipación. Este enfoque permitió una comprensión conceptual crítica sobre aspectos relevantes de la Política Nacional de Asistencia Social y la inserción de la Psicología en esta producción. Se investigó la construcción de la identidad de psicólogos y de ciudadanos usuarios de los Centros de Referencia y Asistencia Social (CRAS), utilizando métodos cualitativos, con énfasis en las entrevistas de Historia de Vida. Entre los fragmentos analizados, se destaca la percepción del psicólogo acerca de su identidad profesional (la angustia y las dificultades sistémicas que obstaculizan sus prácticas emancipatorias). Con relación a los ciudadanos usuarios, se perciben la falta de comprensión sobre el ideario de la política, el no reconocimiento de la Psicología como práctica comunitaria y la conformidad y resignación ante las precariedades que han experimentado.

Abstract The article reflects results of two pieces of scientific initiation research that had the basic scope of the psychosocial theory proposed by professor Ciampa, which conceives identity while human metamorphosis in search for emancipation. This theoretical methodological approach enabled a critical conceptual understanding of relevant aspects of the national policy on social assistance and the insertion of psychology in this production. For this purpose, the identity construction of psychologists and user citizens of the Social Assistance Reference Centers (CRAS) was researched with the use of qualitative methods with emphasis on life story interviews. Among the analyzed fragments, the perception of the psychologist of his/her professional identity (the anguish and systemic problems that hinder his/her emancipatory practices) stands out; as for the user citizens, the observed fragments are lack of comprehension of the policy ideas, the non-recognition of psychology as a community practice, and the conformity and resignation towards the experienced precariousness.

Estandarización de la técnica de PCR-RFLP de la región ITS1, para la caracterización molecular de leishmania (L) infantum, en muestras caninas / Standardization of PCR-RFLP technique of the ITS-1 region for leishmania (L) infantum molecular characterization in canine samples

La leishmaniasis es una enfermedad producida por protozoarios parásitos del género Leishmania. Estos parásitos infectan a hospedadores mamíferos, entre los cuales los perros han sido implicados como reservorios del parásito. Este trabajo planteó estandarizar la técnica de la PCR-RFLP luego de la amplificación de la región ITS1 de Leishmania spp, como herramienta útil para la detección y caracterización molecular. Se utilizaron promastigotes de cultivo y muestras de biopsias procedentes de perros con leishmaniasis visceral previamente diagnosticados en el Centro Antirrábico Nacional. La región ITS1 del ADN genómico nuclear de Leishmania spp. fue amplificada utilizando los cebadores LITSR y L5,8S. La técnica ITS1 PCR-RFLP aplicada, permitió la detección de Leishmania (L) infantum en 10/10 aislados de parásitos mantenidos en medio NNN, en 10/18 muestras de bazo y 10/18 muestras de ganglio linfático poplíteo. Las condiciones óptimas de reacción fueron 0,2 mM de dNTPs, 0,1 pmol de cada cebador y 1U de Taq polimerasa. La sensibilidad de la PCR fue de 3 ng/µL de ADN en aislados de cultivo NNN y 60 ng/µL de ADN en muestras de biopsias, mientras que la especificidad fue de 100% para la detección de Leishmania sp. La enzima de restricción Hae III, determinó fragmentos de 184, 72 y 55 pb., que resultaron específicos para la especie Leishmania (L.) infantum. El marcador utilizado resultó confiable para la detección y caracterización de Leishmania sp. en perros procedentes de zonas endémicas, lo cual podría ser útil para verificar las especies de parásitos circulantes entre los perros.

Leishmaniasis is a disease caused by protozoan parasites of the genus Leishmania. The separasites infect to mammalian hosts, including canines that have been implicated as reservoirs of the parasite. The aim of this research was to standardize the technique of PCR RFLP after amplification of the ITS1 region of Leishmania (Leishmania) infantum, as a useful tool for detection and molecular characterization. Promastigotes from culture and biopsies from dogs with visceral Leishmaniasis previously diagnosed by the Centro Antirrábico Nacional. The ITS1 region of the genomic DNA of Leishmania sp. was amplified using LITSR and L5,8S primers. The technique ITS1 PCR-RFLP applied, allowed the detection of Leishmania (L.) infantum in 10/10 of the isolates from parasites maintained in NNN culture medium, in 10/18 samples from spleen and 10/18 samples from popliteal lymph node. Optimal reaction conditions were 0.2 mM dNTPs, 0.1 pmol of each primer and 1U of Taq polymerase. The sensitivity of PCR was 3 ng/µL DNA in isolates of parasites from NNNculture medium and 60 ng/µL DNA in biopsy samples while the specificity was 100% for the detection of DNA of Leishmania sp. The restriction enzyme Hae III determined fragments of184, 72 and 55 bp., which were specific to Leishmania (L.) infantum. The marker used isreliable for the detection and characterization of Leishmania sp. in dogs from endemic areas, which could be useful to verify the species of parasites circulating among animals.