Phylogenetic relationships of closely-related phlebotomine sand flies (Diptera: Psychodidae) of Nyssomyia genus and Lutzomyia subgenus

BACKGROUND The Nyssomyia genus and Lutzomyia subgenus include medical important species that are Latin American leishmaniases vectors. Little is known about the phylogenetic relationships of closely-related species in each of these taxonomic groups that are morphologically indistinguishable or differentiated by very subtle details. OBJECTIVES We inferred the phylogenetic relationships of closely-related species within both the Nyssomyia genus and the Lutzomyia subgenus using a cytochrome c oxidase subunit I (COI) fragment. METHODS The sampling was carried out from 11 Argentinean localities. For genetic analyses, we used GenBank sequences in addition to our sequences from Argentina. Kimura 2-parameter (K2P) genetic distance and nucleotide divergence (Da) was calculated between closely-related species of Nyssomyia genus, Lutzomyia subgenus and between clades of Lutzomyia longipalpis complex. FINDINGS The K2P and Da values within species of Nyssomyia genus and Lutzomyia subgenus were lower than the divergence detected between clades of Lu. longipalpis complex. The haplotype network analyses within Lutzomyia subgenus showed shared haplotypes between species, contrary to Nyssomyia genus with none haplotype shared. Bayesian inference within Nyssomyia genus presented structuring by species. MAIN CONCLUSIONS This study evidences the phylogenetic proximity among closely-related species within Nyssomyia genus and Lutzomyia subgenus. The COI sequences of Nyssomyia neivai derived from the present study are the first available in GenBank.

An overview of the sandfly fauna (Diptera: Psychodidae) followed by the detection of Leishmania DNA and blood meal identification in the state of Acre, Amazonian Brazil

BACKGROUND In Acre state, Brazil, the dissemination of cutaneous leishmaniasis has increased in recent years, with limited knowledge of the potential Leishmania spp. vectors involved. OBJECTIVES Here, data concerning the sandfly fauna of Brasiléia municipality, Leishmania DNA-detection rates and the identification of blood meal sources of insects captured in 2013-2015 are presented. METHODS Parasite detection in female sandflies was performed individually by multiplex polymerase chain reaction (PCR) (Leishmania kDNA/sandfly cacophony-gene), with the identification of Leishmania spp. by hsp70-PCR and sequencing. The identification of blood gut-content from fed females was performed by cyt b-PCR and sequencing. FINDINGS A total of 4,473 sandflies were captured. A subgroup of 864 non-blood-fed females evaluated for the presence of Leishmania DNA showed 2.9% positivity for Leishmania (Viannia) braziliensis and L. (V.) guyanensis. The identification of blood meal sources was performed in 96 blood-fed females, allowing the identification of 13 vertebrate species. In nine/96 fed females, DNA from L. (V.) shawi, L. (V.) guyanensis, L. (V.) braziliensis and Endotrypanum sp. was detected. MAIN CONCLUSIONS In Brumptomyia sp. and Evandromyia termitophila, the first report of Leishmania DNA-detection is provided in Acre; Nyssomyia shawi is implicated as potential vector of L. (V.) braziliensis and L. (V.) guyanensis for the first time in Brazil.

Ecology of phlebotomine sand flies in a Brazilian area with recent leishmaniasis transmission (Itaúna, in Minas Gerais state)

Abstract INTRODUCTION: Leishmaniasis constitutes a serious but neglected tropical disease. Recently, socio-environmental, biological and physical changes have altered the range of leishmaniasis, causing it to spread into urban areas. In Minas Gerais, the disease is endemic, exhibiting regional differences and reaching urban centers. The purpose of this study was to evaluate entomological aspects related to the ecoepidemiology of leishmaniasis in Itaúna. METHODS: Monthly catches with HP traps were carried out from June 2017 to May 2018, in three ecological areas (urban, rural, and forest). The adaptability of the species to anthropic environments was assessed using the synanthropy index (SI). RESULTS: We collected 1306 specimens of phlebotomine sand flies. Of the species of medical importance, Lutzomyia longipalpis, the vector of Leishmania infantum, represented 90.4% of the specimens identified at species level (n=1260). Nyssomyia whitmani, the vector of Leishmania braziliensis, represented 1.6% of the total. Lu. longipalpis displayed an SI of +95.8, a value that denotes a marked preference for human environments. For Ny. whitmani, the SI was -25, expressing the tendency of this species to occupy uninhabited areas. The population of the three most numerous species captured increased with rain, high temperatures, and high relative humidity. Although captured at low numbers, Ny. whitmani showed a different profile concerning the climate variables analyzed. CONCLUSIONS: Understanding the epidemiology of the disease may assist the health services in formulating effective control measures for improving community health and contributing to the establishment of a dynamic relationship and a global awareness of the health/disease process.

Epidemiological aspects of Leishmaniasis in the Pantanal region of Mato Grosso / Aspectos epidemiológicos das Leishmanioses na região do Pantanal de Mato Grosso

Abstract Leishmaniasis is a disease caused by the protozoan Leishmania parasite that is disseminated by some species of sandflies and hosted by a variety of reservoirs. The objective of the present study was to evaluate the epidemiological situation of leishmaniasis in the municipalities of the Mato Grosso Pantanal. Human data were obtained from the Information System of Notifiable Diseases, and the canine and vector results from the State Department of Health of Mato Grosso. Between 2007 and 2016, 10 cases of visceral leishmaniasis and 499 cases of tegumentary leishmaniasis were identified in the Pantanal region. The surveillance studies regarding the canine reservoir demonstrated that the parasite was present in six of the seven municipalities. Vectors of visceral leishmaniasis were present in five municipalities and vectors of tegumentary leishmaniasis in six. Enhancement of services aimed at controlling this disease is fundamental to prevent an increase in the number of cases in the region.

Resumo As leishmanioses são doenças causadas por protozoários do gênero Leishmania disseminadas por algumas espécies de flebotomíneos e participação de uma variedade de reservatórios. O objetivo do presente estudo foi avaliar a situação epidemiológica das leishmanioses nos municípios abrangentes do Pantanal de Mato Grosso. Os dados humanos foram obtidos do Sistema de Informações de Agravos de Notificação e os resultados caninos e vetoriais junto à Secretaria de Estado de Saúde de Mato Grosso. A região do Pantanal registrou no período de 2007 a 2016, dez casos da forma visceral e 499 casos da forma tegumentar. As ações de vigilância para o reservatório canino demonstraram a circulação do parasito em seis dos sete municípios. A presença dos vetores de leishmaniose visceral ocorreu em cinco municípios e os da leishmaniose tegumentar em seis. O fortalecimento dos serviços voltados para controle desse agravo é fundamental para evitar o aumento no número de casos da região.

Sandflies in an endemic area for Visceral Leishmaniasis in Northeastern Brazil / Flebotomíneos em uma área endêmica para Leishmaniose Visceral no Nordeste do Brasil

Abstract Phlebotomines of the genus Lutzomyia are important vector for species of Leishmania, the etiological agent of leishmaniasis. The aim of this study was to assess the diversity of sandflies in an endemic area for Visceral Leishmaniasis (VL). Sampling was performed in four localities on the coast of the municipality of Goiana, state of Pernambuco, Northeastern Brazil. Specimens were collected monthly on three consecutive nights, from November 2017 to October 2018. CDC light traps were installed at a height of 1.5 m above the ground, in two ecotypes (i.e. tree and animal facility), both in peridomestic areas. A total of 5,205 sandflies were collected, among which the species Lutzomyia longipalpis (99.85%; 5,197/5,205) was the most common, followed by Lutzomyia evandroi (0.13%; 07/5,205) and Lutzomyia whitmani (0.02%; 01/5,205). Specimens of L. longipalpis were collected throughout the study period and most of them were found near to the animal facilities. The findings from this research indicate that vectors for Leishmania infantum are present in the study area throughout the year, along with occurrences of vectors relating to Cutaneous Leishmaniasis (CL). Preventive vector control measures need to be implemented in Goiana to reduce the risk of infection for the human and animal populations.

Resumo Flebotomíneos do gênero Lutzomyia são importantes vetores de espécies de Leishmania, agente etiológico das leishmanioses. O objetivo deste estudo foi avaliar a diversidade de flebotomíneos em área endêmica para Leishmaniose Visceral (LV). As coletas foram realizadas em quatro localidades do litoral do município de Goiana, Pernambuco, Nordeste do Brasil. Os espécimes foram coletados mensalmente em três noites consecutivas de novembro de 2017 a outubro de 2018. Foram instaladas armadilhas luminosas modelo CDC a uma altura de 1,5 m acima do solo em dois ecótopos (ex. árvore e instalações de animais), ambos em áreas peridomiciliares. Um total de 5.205 flebotomíneos foi coletado, sendo Lutzomyia longipalpis (99,85%; 5.197/5.205) a mais comum, seguida por Lutzomyia evandroi (0,13%; 07/5.205) e Lutzomyia whitmani (0,02%; 01/5.205). L. longipalpis foi coletada durante todo o período do estudo. A maioria dos espécimes foi detectada perto das instalações dos animais. Os achados deste estudo indicam a presença de vetores de Leishmania infantum na área avaliada ao longo do ano, bem como a ocorrência de vetores relacionados à Leishmaniose Cutânea (LC). Medidas preventivas de controle vetorial devem ser implementadas em Goiana para reduzir o risco de infecção à população humana e animal.

Sandflies (Diptera: Psychodidae: Phlebotominae), present in an endemic area of cutaneous leishmaniasis in western Boyacá, Colombia / Flebótomos (Diptera: Psychodidae: Phlebotominae), presentes en una zona endémica de leishmaniasis cutánea en el occidente de Boyacá, Colombia

Abstract Introduction: Sandflies are known for having vector species of the tropical disease Leishmaniasis, a disease which is of an endemic nature in Western Boyacá, where the town of Otanche is one of the main source of Cutaneous Leishmaniasis. Objective: To identify the species of sandfly present in an endemic area of cutaneous leishmaniasis in West Boyacá. Methods: The search and collection of sandflies was carried out using CDC gravid traps, over a period of twelve hours (18:00- 06:00). Identification was carried out by revising the genitalia on both male and female samples under a microscope. The distribution took as reference households with a history of people infected with this disease, locating them intra, peri and extra domicile. Results: Were recollected 361 individuals (252 females and 109 male), belonging to 9 genres and 16 species. 60% of all recollected phlebotominae consists of Nyssomyia yuilli and Nyssomyia trapidoi. Other species recollected and relevant, due to vector precedent, are Lutzomyia hartmanni, Psychodopygus panamensis, Lutzomyia gomezi and Psychodopygus carrerai. Conclusion: It was established that, due to its abundance and vector precedent for the country and the area under study, Nyssomyia yuilli and Nyssomyia trapidoi constitute the species of phlebotominae which may be involved in the transmission of cutaneous Leishmaniasis in the region.

Resumen Introducción: Los flebótomos, son conocidos por tener especies vectoras de la enfermedad tropical Leishmaniasis, enfermedad que se presenta con carácter endémico en el occidente del departamento de Boyacá, donde el municipio de Otanche es uno de los principales focos de leishmaniasis cutánea. Objetivo: Identificar las especies de flebótomos presentes en una zona endémica de leishmaniasis cutánea en el occidente del Boyacá. Métodos: La búsqueda y recolección de los flebótomos se realizó con trampas CDC durante doce horas (18:00- 06:00), tomando como referencia viviendas con antecedentes de personas que hubieran tenido la enfermedad, ubicándolas en el intra, peri y extradomicilio. La identificación se realizó por medio de revisión del órgano genital de machos y hembras al microscopio. Resultados: Se colectaron 361 individuos (252 hembras y 109 machos), pertenecientes a 9 géneros y 16 especies, de las cuales, el 60% de toda la flebótomofauna recolectada está representada por Nyssomyia yuilli y Nyssomyia trapidoi. Otras especies colectadas y con importancia por antecedentes vectoriales son Lutzomyia hartmanni, Psychodopygus panamensis, Lutzomyia gomezi y Psychodopygus carrerai. Conclusión: Se estableció, que por sus altas abundancias y por sus antecedentes vectoriales para el país y para la zona de estudio, Nyssomyia yuilli y Nyssomyia trapidoi, constituyen las especies de flebótomos que pueden estar implicadas en la transmisión de leishmaniasis cutánea en la zona de estudio

Molecular detection of Leishmania spp in Lutzomyia longipalpis in the city of Lavras, Minas Gerais, Brazil

Leishmaniasis is a neglected disease that affects a large part of the world population. Knowing the sand fly fauna of a region is of fundamental importance for guiding health surveillance actions related to the prevention and control of leishmaniasis. A total of 86 specimens of sand flies (60 females and 26 males) were collected. Using the classification proposed by Galati (2003), the following species were identified: Lutzomyia longipalpis (Lutz & Neiva, 1912), Migonemyia migonei (França, 1920), Evandromyia cortelezzi (Brethes, 1923), Ev. sallesi (Galvão & Coutinho, 1939), Nyssomyia whitmani (Atunes & Coutinho, 1939), Psathyromyia lutziana (Costa Lima, 1932), Ev. lenti (Mangabeira, 1938), Brumptomyia sp. (França and Parrot, 1921), and Pressatia sp. (Mangabeira, 1942). Using PCR with internal transcribed spacer target to identify infected sand flies, five Lu. longipalpis females were infected with Leishmania spp. Despite the small number of specimens collected, considerable species diversity was found in the study area.

Lutzomyia longipalpis (Diptera: Psychodidae) Argentina-Bolivia border: new report and genetic diversity

American visceral leishmaniasis (AVL) has two main scenarios of transmission as follows: scattered cases in rural areas and urban outbreaks. Urban AVL is in active dispersion from the northeastern border of Argentina-Paraguay-Brazil to the South. The presence of Lutzomyia longipalpis was initially reported in urban environments in the northwestern border of the country. The presence of Lu. longipalpis, environmental variables associated with its distribution, and its genetic diversity were assessed in Salvador Mazza, Argentina, on the border with Bolivia. The genetic analysis showed high haplotype diversity, low nucleotide diversity, and low nucleotide polymorphism index. We discuss the hypothesis of an expanding urban population with introgressive hybridisation of older haplogroups found in their path in natural forest or rural environments, acquiring a new adaptability to urban environments, and the possibility of changes in vector capacity.

Molecular validation of anthropophilic Phlebotominae sandflies (Diptera: Psychodidae) in Central Panama

Six Phlebotominae sand fly species are incriminated as biological vectors of human pathogens in Panama, but molecular corroboration is still needed. We aim at confirming the identity of Phlebotominae species documented as anthropophilic in Panama. Adult sandflies were collected from August 2010 to February 2012 in Central Panama using CDC light traps. Species confirmation was accomplished through molecular barcodes and allied sequences from GenBank. A total of 53,366 sand fly specimens representing 18 species were collected. Five species were validated molecularly as single phylogenetic clusters, but Psychodopygus thula depicted two genetically divergent lineages, which may be indicative of cryptic speciation.

Nocturnal activity rhythm of nyssomyia intermedia (diptera: psychodidae: phlebotominae) in campus fiocruz mata atlântica, an american cutaneous leishmaniasis transmission area, Rio de Janeiro, Brazil

Abstract INTRODUCTION: Nyssomyia intermedia is an important vector of American cutaneous leishmaniasis. METHODS: Sand flies were captured in a Shannon trap and after landing on human collectors and retrieved monthly. Nocturnal activity was estimated using a log-linear model. RESULTS: Peak sand fly collection in the Shannon trap occurred in summer and winter, and peak fly collection on humans occurred in spring and summer. CONCLUSIONS: Ny. intermedia was captured in both the hottest and coldest months, indicating its adaptability to different seasons and the possibility of disease transmission at any time of the year in Campus Fiocruz Mata Atlântica, Brazil.

A new species of phlebotomine sandfly, Pintomyia (Pifanomyia) duckei (Diptera: Psychodidae) from Manaus, State of Amazonas, Brazil / Uma nova espécie de flebotomíneo, Pintomyia (Pifanomyia) duckei (Diptera: Psychodidae) de Manaus, Estado do Amazonas, Brasil

Pintomyia (Pifanomyia) duckei Oliveira, Alencar and Freitas sp. n. from Adolpho Ducke Reserve, Manaus, Amazonas state, Brazil, is described and illustrated based on three male specimens, the holotype and two paratypes. The paratypes have abnormalities in the external genitalia, one with asymmetric bilateral anomaly (different anomalies in each of certain paired structures) and the other with unilateral anomaly (anomaly in one of the paired structures). The new species probably belongs to the Serrana Series, integrating the list of the species whose males possess gonostylus with two stout spines (one apical and other subapical), as well as a small and very thin spine on the inner surface of the basal half.

Pintomyia (Pifanomyia) duckei Oliveira, Alencar e Freitas sp. n. da Reserva Florestal Adolpho Ducke, Manaus, Estado do Amazona, Brasil, é descrita e ilustrada com base em três espécimes machos, o holótipo e dois parátipos. Os parátipos possuem anomalias na genitália externa, sendo um com anomalia bilateral assimétrica (anomalias diferentes em cada uma das estruturas pareadas) e outro com anomalia unilateral (anomalia em uma das estruturas pareadas). A nova espécie provavelmente pertence à Série Serrana, integrando a lista das espécies, cujos machos possuem gonóstilo com dois espinhos robustos (um apical e outro subapical), bem como um espinho delgado e muito curto na superfície interna da metade basal.

Lutzomyia umbratilis from an area south of the Negro River is refractory to in vitro interaction with Leishmania guyanensis

BACKGROUND Lutzomyia umbratilis, the vector for Leishmania guyanensis in northern South America, has been found naturally infected with L. guyanensis only in areas north of the Negro and Amazon rivers. While populations of this sand fly species are also found in areas south of these rivers, these populations have never been reported to be infected and/or transmitting L. guyanensis. However, no studies on the corresponding host-parasite interactions are available. OBJECTIVES This study evaluated the interaction between Lu. guyanensis promastigotes and field-collected Lu. umbratilis sand flies from Rio Preto da Eva and Manacapuru, which are located to the north and south, respectively, of the Negro River. METHODS Procyclic and metacyclic attachment was quantified using an in vitro system. FINDINGS Low attachment of parasites to the midguts of insects collected from Manacapuru was detected. Conversely, greater binding of metacyclic parasites was observed in the midguts of insects collected from Rio Preto da Eva, and this attachment was more pronounced than that observed for procyclics (p < 0.03). MAIN CONCLUSIONS The Lu. umbratilis population from an area south of the Negro River has lower in vitro interaction with L. guyanensis. The higher attachment of L. guyanensis to midguts of insects from Rio Preto da Eva may suggest better vector competence. These findings are in accordance with previously reported epidemiological information of American cutaneous leishmaniasis (ACL) transmission in the Amazon.

Taxonomia integrativa de espécies, com fêmeas morfologicamente similares, do gênero Psychodopygus (Diptera, Psychodidae), Série Chagasi, registradas no Brasil / Integrative taxonomy of morphologically indistinguishable species of the genus Psychodopygus (Diptera, Psychodidae), Chagasi series, registered in Brazil

Introdução. A identificação dos flebotomíneos baseia-se principalmente na morfologia do adulto, o que pode ser problemático quando as espécies são morfologicamente muito semelhantes. Psychodopygus é um gênero de flebotomíneos de grande interesse em saúde pública devido ao papel de algumas espécies na veiculação de Leishmania spp. no Brasil. No entanto, este gênero inclui espécies com fêmeas morfologicamente indistinguíveis que pertencem à Série Chagasi, sendo elas: P. chagasi, P. complexus, P. squamiventris maripaensis, P. squamiventris squamiventris e P. wellcomei. Objetivos. Investigar a possibilidade de distinguir essas espécies por meio de análises morfométrica e molecular, além de produzir uma distribuição geográfica atualizada para o grupo analisando a probabilidade de ocorrência das espécies através da análise de modelagem de nicho ecológico. Material e Métodos. Foi realizada a análise discriminante na morfometria geométrica (cabeça e asa) e linear, morfologia (usando microscopia óptica e eletrônica de varredura) e a análise do citocromo c oxidase subunidade 1 (COI), avaliando-se um total de 752 espécimes (460 fêmeas e 292 machos) dos seguintes estados Amapá, Amazonas, Ceará, Mato Grosso, Pará, Rondônia, Roraima e Tocantins. Mapas de distribuição foram produzidos através de dados obtidos do material analisado e de revisão bibliográfica. Resultados. A análise discriminante usando caracteres morfométricos lineares mostrou-se capaz de diferenciar todas as espécies, exceto P. complexus, que apresentou 2,2% de erro de identificação. A morfometria geométrica das asas foi incapaz de separar completamente as espécies através da conformação, mas o tamanho do centróide dos espécimes fêmeas falhou apenas em distinguir P. complexus de P. s. maripaensis. Por outro lado, a morfometria geométrica das cabeças foi capaz de distinguir todas as espécies com grande eficiência ao usar tanto a forma como o tamanho do centróide. A análise morfológica revelou que a coloração torácica, principalmente do pronoto e do pós-noto, pode ser usada para separar as cinco espécies em três grupos: P. chagasi, P. wellcomei / P. complexus e P. s. mariapaensis / P. s. squamiventris. Os resultados da análise de DNA Barcoding, mostraram um agrupamento semelhante ao observado na morfologia; embora os espécimes de P. wellcomei do estado do Ceará mostrem uma grande distância genética da população do estado do Pará, evidenciando que essa espécie possa representar um complexo. Quanto à microscopia eletrônica de varredura, foram avaliadas detalhadamente as estruturas das antenas, tórax e genitália masculina. Salientamos que no anepímero (tórax) foi observada uma escama tipo \"raquete\" modificada apenas em Psychodopygus s. squamiventris. A revisão da distribuição geográfica mostrou que as espécies possuem uma distribuição cis-andina, ocorrendo principalmente no bioma Amazônico. A nítida separação de algumas espécies pelo rio Amazonas, sugere que o surgimento do grupo ocorreu no período que se estende da orogênese dos Andes até a formação deste rio. Conclusões. O estudo possibilitou diferenciar completamente as fêmeas das cinco espécies da Série Chagasi utilizando o conjunto de dados obtidos por morfometria linear e geométrica e análises morfológicas e também apresentar novos caracteres morfológicos e padrões distribucionais que facilitarão a identificação de machos e fêmeas dessas espécies

Introduction. The identification of sand flies is mainly based on adult morphology, which can be problematic when species are morphologically very similar. Psychodopygus is one of the sand fly genera of great interest in public health, due to the role of some species in the transmission of Leishmania spp. in Brazil. However, this genus includes species with morphologically indistinguishable females that belong to the Chagasi series, which includes: P. chagasi, P. complexus, P. squamiventris maripaensis, P. squamiventris squamiventris and P. wellcomei. Objectives. To investigate the possibility of distinguishing among these species by means of morphometric and molecular analyses in addition to producing an updated geographical distribution for the group, analyzing the probability of the occurrence of the species by the analysis of ecological niche modeling. Material and methods. The analyses of the cytochrome c oxidase subunit 1 (COI), geometrical (head and wing) and of linear morphometry and morphology (using optical microscopy and scanning electron microscopy) were carried out using a total of 752 specimens (460 females and 292 males) from the following states: Amapá, Amazonas, Ceará, Mato Grosso, Pará, Rondônia, Roraima e Tocantins. Distribution maps were produced on the basis of data obtained from the material analyzed and a bibliographical review. Results. The discriminant analysis using linear morphometric characters was able to differentiate among all the species, except for P. complexus, which presented a 2.2% error of identification. The geometric morphometry of the wings was unable to completely separate the species by means of the shape analyses, but the centroid size of the female specimens only failed to distinguish P. complexus from P. s. maripaensis. Otherwise, the geometric morphometry of the heads was sufficient to distinguish all the species with great efficiency, when using both the head-shape and the centroid size. The morphological analysis revealed that the thoracic coloration, mainly of the pronotum and the post-notum, can be used to separate the five species into three groups: P. chagasi, P. wellcomei / P. complexus, P. s. mariapaensis / P. s. squamiventris. The results of the Barcoding DNA analyses showed a cluster similar to that observed in the morphology; however, P. wellcomei specimens from the Ceará population showed a great genetic distance from the population of Pará, evidencing that this species may represent a complex. As for the scanning electron microscopy, the structures of the antennae, thorax and male genitalia were evaluated in detail. In the anepimerum (thorax) a modified \"racket\"-type scale was observed only in Psychodopygus s. squamiventris. The review of the geographical distribution showed that the species have a cis-Andean distribution, occurring mainly in the Amazonian biome. The separation of some species from the others by the Amazon river suggests that the appearance of the Chagasi series occurred in the period from the orogenesis of the Andes to the formation of this river. Conclusions. The results clearly differentiate the females of the five species of the Chagasi series using the data set of linear and geometric morphometry and morphological analyses, providing new morphological and distributional data that will facilitate the identification of the males and females of this group

The role of gallery forests in maintaining Phlebotominae populations: potential Leishmania spp. vectors in the Brazilian savanna

BACKGROUND Knowledge on synanthropic phlebotomines and their natural infection by Leishmania is necessary for the identification of potential areas for leishmaniasis occurrence. OBJECTIVE To analyse the occurrence of Phlebotominae in gallery forests and household units (HUs) in the city of Palmas and to determine the rate of natural infection by trypanosomatids. METHODS Gallery forests and adjacent household areas were sampled on July (dry season) and November (rainy season) in 2014. The total sampling effort was 960 HP light traps and eight Shannon traps. Trypanosomatids were detected in Phlebotominae females through the amplification of the SSU rDNA region, and the positive samples were used in ITS1-PCR. Trypanosomatid species were identified using sequencing. FINDINGS A total of 1,527 sand flies representing 30 species were captured in which 949 (28 spp.) and 578 (22 spp.) were registered in July and November, respectively. In July, more specimens were captured in the gallery forests than in the HUs, and Nyssomyia whitmani was particularly frequent. In November, most of the specimens were found in the HUs, and again, Ny. whitmani was the predominant species. Lutzomyia longipalpis was commonly found in domestic areas, while Bichromomyia flaviscutellata was most frequent in gallery forests. Molecular analysis of 154 pools of females (752 specimens) identified Leishmania amazonensis, L. infantum, and Crithidia fasciculata in Ny. whitmani, as well as L. amazonensis in Lu. longipalpis, Trypanosoma sp. and L. amazonensis in Pintomyia christenseni, and L. amazonensis in both Psathyromyia hermanlenti and Evandromyia walkeri. MAIN CONCLUSIONS These results show the importance of gallery forests in maintaining Phlebotominae populations in the dry month, as well as their frequent occurrence in household units in the rainy month. This is the first study to identify Leishmania, Trypanosoma, and Crithidia species in Phlebotominae collected in Palmas, Tocantins, Brazil.

Environmental suitability for Lutzomyia longipalpis in a subtropical city with a recently established visceral leishmaniasis transmission cycle, Argentina

BACKGROUND Visceral leishmaniasis (VL) is an endemic disease in northeastern Argentina including the Corrientes province, where the presence of the vector and canine cases of VL were recently confirmed in December 2008. OBJECTIVES The objective of this study was to assess the modelling of micro- and macro-habitat variables to evaluate the urban environmental suitability for the spatial distribution of Lutzomyia longipalpis presence and abundance in an urban scenario. METHODS Sampling of 45 sites distributed throughout Corrientes city (Argentina) was carried out using REDILA-BL minilight traps in December 2013. The sampled specimens were identified according to methods described by Galati (2003). The analysis of variables derived from the processing of satellite images (macro-habitat variables) and from the entomological sampling and surveys (micro-habitat variables) was performed using the statistical software R. Three generalised linear models were constructed composed of micro- and macro-habitat variables to explain the spatial distribution of the abundance of Lu. longipalpis and one composed of micro-habitat variables to explain the occurrence of the vector. FINDINGS A total of 609 phlebotominae belonging to five species were collected, of which 56% were Lu. longipalpis. In addition, the presence of Nyssomyia neivai and Migonemya migonei, which are vectors of tegumentary leishmaniasis, were also documented and represented 34.81% and 6.74% of the collections, respectively. The explanatory variable normalised difference vegetation index (NDVI) described the abundance distribution, whereas the presence of farmyard animals was important for explaining both the abundance and the occurrence of the vector. MAIN CONCLUSIONS The results contribute to the identification of variables that can be used to establish priority areas for entomological surveillance and provide an efficient transfer tool for the control and prevention of vector-borne diseases.

Especies de flebotomíneos (Diptera: Psychodidae) recolectados en reservas naturales de las regiones del Darién y del Pacífico en Colombia / Species of sand flies (Diptera: Psychodidae) collected from natural reserves in the Pacific and Darien regions of Colombia

Resumen Introducción. Los departamentos de Chocó y Antioquia en Colombia presentan condiciones climáticas y de vegetación que favorecen el establecimiento de especies de vectores del género Lutzomyia y la transmisión de Leishmania spp. a poblaciones humanas que ingresan a ambientes selváticos conservados. Objetivo. Reportar las especies de flebotomíneos presentes en tres reservas naturales de las regiones del Darién y del Pacífico en Colombia. Materiales y métodos. Los flebotomíneos se recolectaron en las reservas naturales El Aguacate (Acandí, Chocó), Nabugá (Bahía Solano, Chocó) y Tulenapa (Carepa, Antioquia). La recolección se hizo con trampas de luz CDC, mediante búsqueda activa en sitios de reposo y con trampas Shannon. La determinación taxonómica de especies se basó en las claves taxonómicas. En algunas especies de interés taxonómico, se evaluó el uso de secuencias parciales de la región 5' del gen COI. Resultados. Se recolectaron 611 flebotomíneos adultos: 531 en Acandí, 45 en Carepa y 35 en Bahía Solano. Se identificaron 17 especies del género Lutzomyia, tres del género Brumptomyia y una del género Warileya. Las distancias genéticas (K2P) y los soportes de agrupación (>99 %) en el dendrograma de neighbor joining correspondieron a la mayoría de unidades taxonómicas operacionales moleculares (Molecular Operational Taxonomic Units, MOTU) establecidas para el grupo Aragaoi y confirmaron claramente la identidad de Lu. coutinhoi. Conclusión. Se identificaron especies que tienen importancia en la transmisión de la leishmaniasis en Acandí, Bahía Solano y Carepa. Se confirmó la presencia de Lu. coutinhoi en Colombia.

Abstract Introduction: The departments of Chocó and Antioquia in Colombia show climatic and vegetation conditions favoring the establishment of vector species of the genus Lutzomyia and the transmission of Leishmania spp. to human populations entering conserved forest environments. Objective: To report the species of Phlebotomine sandflies present in three natural reserves in the Darien and Pacific regions of Colombia. Materials and methods: Sand flies were collected specifically in the natural reserves El Aguacate (Acandí, Chocó), Nabugá (Bahía Solano, Chocó) and Tulenapa (Carepa, Antioquia). Sand flies were collected with CDC light traps, active search in resting places and Shannon traps. The taxonomic determination of species was based on taxonomic keys. For some species of taxonomic interest, we evaluated the partial sequences of the 5' region of COI gene. Results: A total of 611 adult sand flies were collected: 531 in Acandí, 45 in Carepa and 35 in Bahía Solano. Seventeen species of the genus Lutzomyia, three of the genus Brumptomyia and one of the genus Warileya were identified. The genetic distances (K2P) and grouping supported (>99%) in the neighbor joining dendrogram were consistent for most established molecular operational taxonomic units (MOTU) of the Aragaoi group and clearly confirmed the identity of Lu. coutinhoi. Conclusion: Species that have importance in the transmission of leishmaniasis in Acandí, Bahía Solano and Carepa were identified. The presence of Lu. coutinhoi was confirmed and consolidated in Colombia.

Seasonality of sand flies (Diptera: Psychodidae) and Leishmania DNA detection in vector species in an area with endemic visceral leishmaniasis

BACKGROUND Leishmaniases are a serious health problem in southeast Brazil, including the city of Belo Horizonte (BH), Minas Gerais state (MG), where there are high rates of incidence and mortality due to visceral leishmaniases. BH is divided into nine sanitary districts (SD) of which one, the Venda Nova SD, was selected for this study because it has high rates of positivity for canine leishmaniasis and high incidence of human leishmaniasis. OBJECTIVES This study aimed to survey the sand fly fauna in Venda Nova SD from August 2011 to July 2013 and perform a descriptive analysis of the vector population. METHODS The sampling was carried out using automatic HP light traps at all covered areas of the Venda Nova SD, in a total of eighteen light traps. Sampled specimens were identified following Galati (2003), and females were submitted to molecular techniques for the detection and identification of Leishmania DNA. A simple environmental description was done for it area and Kernel estimation was used to infer vector density for each study site. FINDINGS A total of 2,427 sand fly specimens belonging to eight species and five genera were collected of which 95.3% were Lutzomyia longipalpis. The seasonal variation curve was delineated by this species. Lu. longipalpis was the most abundant at all collection points and in all months of the study, and exhibited a natural infection rate of 1.01% for Leishmania infantum and 1.77% for Leishmania braziliensis. MAIN CONCLUSIONS The results show the presence and adaptation of Lu. longipalpis to the anthropic environment of BH and reinforces its role as the main vector of L. infantum in the region.

The current status of the Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) species complex

Lutzomyia longipalpis s.l. is a complex of sibling species and is the principal vector of American visceral leishmaniasis. The present review summarises the diversity of efforts that have been undertaken to elucidate the number of unnamed species in this species complex and the phylogenetic relationships among them. A wide variety of evidence, including chemical, behavioral and molecular traits, suggests very recent speciation events and complex population structure in this group. Although significant advances have been achieved to date, differential vector capacity and the correlation between structure of parasite and vector populations have yet to be elucidated. Furthermore, increased knowledge about recent epidemiological changes, such as urbanisation, is essential for pursuing effective strategies for sandfly control in the New World.

Entering and exiting behaviour of the phlebotomine sand fly Lutzomyia longiflocosa (Diptera: Psychodidae) in rural houses of the sub-Andean region of Colombia

The present study identified the entering and exiting sites for Lutzomyia longiflocosa in rural houses of the sub-Andean region in Colombia. Entering sites were identified with sticky traps set up outside the bedrooms, around the eave openings, and with cage traps enclosing the slits in the doors and windows inside the bedrooms. Exiting sites were identified by releasing groups of females indoors. These females were blood fed and marked with fluorescent powders. Females were recaptured with the trap placement described above but set up on the opposite sides of the openings. In the entering experiment, a significantly higher number of females were captured in the sticky traps at the zone nearest the eave openings (n = 142) than those captured in the other zones of the trap (n = 52); similarly, a higher number of females were captured on the front side of the house (n = 105) than at the rear side (n = 37). Only two females were collected in the cage trap. In the exiting experiment, at the ceiling, the highest percentage (86.2%) of females was recaptured with sticky traps nearest the eave openings and on the front side of the house (70.0%). Seven females were collected in the cage trap. Lu. longiflocosa entered and exited houses primarily through the eave openings in a non-random pattern in relation to the sides of the house.

First description of Leishmania (Viannia) infection in Evandromyia saulensis, Pressatia sp. and Trichophoromyia auraensis (Psychodidae: Phlebotominae) in a transmission area of cutaneous leishmaniasis in Acre state, Amazon Basin, Brazil

Studies on the sandfly fauna to evaluate natural infection indexes are still limited in the Brazilian Amazon, a region with an increasing incidence of cutaneous leishmaniasis. Here, by using a multiplex polymerase chain reaction directed to Leishmania kDNA and hybridisation, we were able to identify L. (Viannia) subgenus in 12 out of 173 sandflies captured in the municipality of Rio Branco, Acre state, revealing a positivity of 6.94%. By sequencing the Leishmania 234 bp-hsp70 amplified products from positive samples, infection by L. (V.) braziliensis was confirmed in five sandflies: one Evandromyia saulensis, three Trichophoromyia auraensis and one Pressatia sp. The finding of L. (Viannia) DNA in two Ev. saulensis corresponds to the first record of possible infection associated with this sandfly. Moreover, our study reveals for the first time in Brazil, Th. auraensis and Pressatia sp. infected by L. (Viannia) parasites.