Identification of blood meal sources of Lutzomyia longipalpis using polymerase chain reaction-restriction fragment length polymorphism analysis of the cytochrome B gene

An analysis of the dietary content of haematophagous insects can provide important information about the transmission networks of certain zoonoses. The present study evaluated the potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mitochondrial cytochrome B (cytb) gene to differentiate between vertebrate species that were identified as possible sources of sandfly meals. The complete cytb gene sequences of 11 vertebrate species available in the National Center for Biotechnology Information database were digested with Aci I, Alu I, Hae III and Rsa I restriction enzymes in silico using Restriction Mapper software. The cytb gene fragment (358 bp) was amplified from tissue samples of vertebrate species and the dietary contents of sandflies and digested with restriction enzymes. Vertebrate species presented a restriction fragment profile that differed from that of other species, with the exception of Canis familiaris and Cerdocyon thous. The 358 bp fragment was identified in 76 sandflies. Of these, 10 were evaluated using the restriction enzymes and the food sources were predicted for four: Homo sapiens (1), Bos taurus (1) and Equus caballus (2). Thus, the PCR-RFLP technique could be a potential method for identifying the food sources of arthropods. However, some points must be clarified regarding the applicability of the method, such as the extent of DNA degradation through intestinal digestion, the potential for multiple sources of blood meals and the need for greater knowledge regarding intraspecific variations in mtDNA.

A comparison of molecular markers to detect Lutzomyia longipalpis naturally infected with Leishmania (Leishmania) infantum

The aim of the present study was to detect natural infection by Leishmania (Leishmania) infantum in Lutzomyia longipalpis captured in Barcarena, state of Pará, Brazil, through the use of three primer sets. With this approach, it is unnecessary to previously dissect the sandfly specimens. DNA of 280 Lu. longipalpis female specimens were extracted from the whole insects. PCR primers for kinetoplast minicircle DNA (kDNA), the mini-exon gene and the small subunit ribosomal RNA (SSU-rRNA) gene of Leishmania were used, generating fragments of 400 bp, 780 bp and 603 bp, respectively. Infection by the parasite was found with the kDNA primer in 8.6% of the cases, with the mini-exon gene primer in 7.1% of the cases and with the SSU-rRNA gene primer in 5.3% of the cases. These data show the importance of polymerase chain reaction as a tool for investigating the molecular epidemiology of visceral leishmaniasis by estimating the risk of disease transmission in endemic areas, with the kDNA primer representing the most reliable marker for the parasite.

Incipient colonisation of Lutzomyia longipalpis in the city of Resistencia, province of Chaco, Argentina (2010-2012)

Lutzomyia longipalpis was recorded for the first time in Argentina in 2004, in the province of Formosa. In the following years, the vector spread to the south and west in the country and was recorded in the province of Chaco in 2010. From November 2010-May 2012, captures of Phlebotominae were made in the city of Resistencia and its surroundings, to monitor the spread and possible colonisation of Lu. longipalpis in the province of Chaco. In this monitoring, Lu. longipalpis was absent in urban sampling sites and its presence was restricted to Barrio de los Pescadores. This suggests that the incipient colonisation observed in 2010 was not followed by continuous installation of vector populations and expansion of their spatial distribution as in other urban centres of Argentina.

Phlebotomine fauna, natural infection rate and feeding habits of Lutzomyia cruzi in Jaciara, state of Mato Grosso, Brazil

Visceral leishmaniasis (VL) in Brazil is transmitted by the phlebotomine Lutzomyia longipalpis and in some midwestern regions by Lutzomyia cruzi. Studies of the phlebotomine fauna, feeding habits and natural infection rate by Leishmania contribute to increased understanding of the epidemiological chain of leishmaniases and their vectorial capacity. Collections were performed in Jaciara, state of Mato Grosso from 2010-2013, during which time 2,011 phlebotomines (23 species) were captured (68.70% Lu. cruzi and 20.52% Lutzomyia whitmani). Lu. cruzi females were identified by observing the shapes of the cibarium (a portion of the mouthpart) and spermatheca, from which samples were obtained for polymerase chain reaction to determine the rates of natural infection. Engorged phlebotomines were assessed to identify the blood-meal host by ELISA. A moderate correlation was discovered between the number of Lu. cruzi and the temperature and the minimum rate of infection was 6.10%. Twenty-two females were reactive to the antisera of bird (28%), dog (3.30%) and skunk (1.60%). We conclude that Lu. cruzi and Lu. whitmani have adapted to the urban environment in this region and that Lu. cruzi is the most likely vector of VL in Jaciara. Moreover, maintenance of Leishmania in the environment is likely aided by the presence of birds and domestic and synanthropic animals.

Lutzomyia longipalpis urbanisation and control

Since the description of Lutzomyia longipalpis by Lutz and Neiva more than 100 years ago, much has been written in the scientific literature about this phlebotomine species. Soares and Turco (2003) and Lainson and Rangel (2005) have written extensive reviews focused on vector-host-parasite interactions and American visceral leishmaniasis ecology. However, during the last two decades, the success of Lu. longipalpis in colonising urban environments and its simultaneous geographical spreading have led to new theoretical and operational questions. Therefore, this review updates the general information about this species and notes the more challenging topics regarding the new scenario of urbanisation-spreading and its control in America. Here, we summarise the literature on these issues and the remaining unsolved questions, which pose recommendations for operational research.

Abundance of Lutzomyia longipalpis in urban households as risk factor of transmission of visceral leishmaniasis

Urban occurrence of human and canine visceral leishmaniasis (VL) is linked to households with characteristics conducive to the presence of sand flies. This study proposes an ad hoc classification of households according to the environmental characteristics of receptivity to phlebotominae and an entomological study to validate the proposal. Here we describe the phlebotominae population found in intra- and peridomiciliary environments and analyse the spatiotemporal distribution of the VL vector Lutzomyia longipalpis of households receptive to VL. In the region, 153 households were classified into levels of receptivity to VL followed by entomological surveys in 40 of those properties. Kruskal-Wallis verified the relationship between the households’ classification and sand fly abundance and Kernel analysis evaluated L. longipalpis spatial distribution: of the 740 sand flies were captured, 91% were L. longipalpis; 82% were found peridomiciliary whilst the remaining 18% were found intradomiciliary. No statistically significant association was found between sandflies and households levels. L. longipalpis counts were concentrated in areas of high vulnerability and some specific households were responsible for the persistence of the infestation. L. longipalpis prevails over other sand fly species for urban VL transmission. The entomological study may help target the surveillance and vector control strategies to domiciles initiating and/or maintaining VL outbreaks.

Description of Lutzomyia chotensis, a new species of Peruvian Phlebotominae (Diptera, Psychodidae)

Lutzomyia chotensis sp. nov. (Diptera, Psychodidae, Phlebotominae) from Cajamarca Department, Peru, is described and its taxonomic position discussed.

Description of Lutzomyia (Helcocyrtomyia) herreri sp. nov. (Diptera, Psychodidae, Phlebotominae) from the South Peruvian Andes

A new species of phlebotomine, Lutzomyia (Helcocyrtomyia) herreri sp. nov., belonging to the series osornoi, from the Department of Puno, Peru, is described. Identification keys for both sexes of the species of this series are presented.

Descripción del ARN de transferencia mitocondrial para Serina (UCN) de Lutzomyia columbiana (Diptera, Psychodidae)

Lutzomyia columbiana es un flebotomíneo considerado como vector sospechoso de Leishmania mexicana y Leishmania braziliensis en Colombia. Este insecto pertenece al grupo verrucarum, que incluye algunos taxones isomórficos, lo que ha estimulado la búsqueda de marcadores moleculares que permitan, además de diferenciar las especies, estudiar sus relaciones de parentesco. En este artículo se describe por primera vez la estructura putativa del ARN de transferencia mitocondrial para serina que reconoce el codón UCN (ARNtSer) de Lu. columbiana. El ADN genómico fue extraído, amplificado y secuenciado a partir de seis especímenes colectados con cebo humano. La estructura secundaria del ARNtSer fue inferida con el programa tRNAscan-SE 1.21. El gen ARNts consistió de 67 pares de bases (pb), encontrándose un solo haplotipo en los seis individuos secuenciados. El ARNtSer de Lu. columbiana mostró 7 apareamientos intracatenarios en el brazo aceptor del aminoácido, 3 en el brazo dihidrouridina (DHU), 5 en el brazo del anticodón y 5 en el brazo ribotimidina-pseudouridina-citosina (TøC). El tamaño de las lupas correspondió a 5 nucleótidos en la DHU, 7 en la anticodón, 4 en la variable y 7 en la TøC. Lu. columbiana se distingue del resto de especies de Lutzomyia y Phlebotomus secuenciadas a la fecha por la presencia de una guanina en la posición nucleotídica 64, que produce un apareamiento no canónico tipo uracilo-guanina en el brazo aceptor. Se necesitan más estudios para confirmar la utilidad del ARNtSer como marcador molecular para la discriminación de especies de flebotomíneos.

Flebotomíneos encontrados em galinheiros experimentais nos Estados de São Paulo e Minas Gerais (Brasil) e algumas observações ecológicas

Relata-se o encontro de várias espécies de Phlebotominae em galinheiros experimentais distribuídos nos municípios de Guaíra, Pariquera-Açu, Taquarituba, no Estado de São Paulo, e Frutal, Sacramento, no Estado de Minas Gerais, Brasil. Assinala-se a colonização de Lutzomyia migonei (França, 1920), alguns dados sobre Psychodopygus intermedius (Lutz & Neiva, 1912), bem como, novos encontros.

Foco de leishmaniose tegumentar americana nos municípios de Viana e Cariacica, Estado do Espirito Santo, Brasil

Descreve-se um foco de leishmaniose tegumentar americana abrangendo quatro localidades dos municípios de Viana e Cariacica situados próximo ao litoral do Estado do Espírito Santo, Brasil. Foram diagnosticados 71 casos no período de maio de 1978 a dezembro de 1982. A confirmação laboratorial foi obtida pela intradermorreação de Montenegro em 66 pacientes e pelo encontro do parasita em 15. A faixa etária mais acometida foi a de 0-10 anos. Amostras do parasita isoladas de 3 pacientes e de 27 cães foram classificadas como Leishmania braziliensis braziliensis. De apenas dois Proechimys iheringi, entre 257 mamíferos silvestres estudados, isolou-se Leishmania, que foi identificada como Leishmania mexicana aristedesi. As espécies de flebotomíneos mais numerosas (Lutzomyia intermedia, L. fischeri e L. migonei) apresentaram um elevado grau de adaptação aos ambientes intra e peri-domibiliar. Levantou-se a hipótese da transmissão estar ocorrendo no intra e peri-domicílio.

Ecological aspects of American cutaneous leishmaniasis: 4. Observations on the endophilic behavior of the sandfly and the vectorial role of Psychodopygus intermedius in the Ribeira Valley region of the S.Paulo State, Brazil

The invasive tendency of Psychodopygus intermedius in the home environment, observed initially by Forattini et al. (1976), has now been confirmed by the demonstration of its high endophilic ability and by the use of human residences for shelter. Populations such as Lutzomyia migonei and Pintomyia fischeri were also present in that environment, though their low densities registered during this investigation could be an indication of their poor ability to overcome the barriers raised by the artificial environment. An objective epidemiological analysis based on the variables here given showed that human infection takes place in the extraforest environment, and the principal vectorial function falls, without doubt, on P. intermedius.

Observações sobre a leishmaniose tegumentar, cinco anos após profilaxia

Em 1974 foram diagnosticados vários casos de leishmaniose tegumentar americana, no Município do Rio de Janeiro, RJ (Brasil). Naquela ocasião, foram usadas como medidas profiláticas o tratamento dos doentes e a dedetização dos domicílios e anexos, resultando na interrupção da transmissão. Com o aparecimento de novos casos, a partir de 1979, foram reestudadas algumas características de uma das localidades (Camorim) do mesmo município. De junho de 1979 a agosto de 1981, foram comprovados nessa localidade 19 casos novos de leishmaniose tegumentar americana (18 humanos e um em cão). Coletaram-se 1.149 flebótomos (11,4% fêmeas e 88,6% machos) com predomínio de Lutzomyia intermedia (95%). De acordo com o estudo dos casos, pressupõe-se que a transmissão tenha ocorrido no domicílio ou peridomicílio. Acredita-se, pois, que na área estudada, as medidas de controle, adotadas em 1974, foram eficientes por um período de aproximadamente quatro anos.

Nota sobre encontro de Phlebotominae (Diptera: Psychodidae) no litoral sul do Estado de São Paulo, Brasil

Durante as realizações de dois estudos sobre mosquito Culicidae, empregando-se armadilhas luminosas tipo CDC e Shanon, além de isca humana, foram capturados cinco espécies de Phlebotominae, entre as quais se encontravam Lutzomyia intermedia e L. flaviscutellata. As localidades investigadas eram alagadiças e raros casos humanos de leishmaniose tegumentar foram registrados entre os moradores locais. As duas situações sugerem ser estes casos autóctones.