Natural sandfly infection by Leishmania spp. in an area endemic for American cutaneous leishmaniasis was analyzed using multiplex polymerase chain reaction (PCR). The sandflies were captured using Falcão light traps in an endemic area of the municipality of Doutor Camargo during March, April, and June 2008. In total, 1803 females were analyzed; 1755 were Nyssomyia neivai (Pinto) and 48 were Nyssomyia whitmani (Antunes and Coutinho). Multiplex PCR analyses using MP3H-MP1L and 5Llcac-3Llcac primers showed the presence of Leishmania (Viannia) spp. in 4/181 pools of sandflies, all Ny. neivai, that is, a minimal infection rate of 0.22%. This study showed, for the first time, the presence of DNA of Leishmania (Viannia) spp. in Ny. neivai. This suggests the existence of natural infection by Leishmania (Viannia) spp. in Ny. neivai in the state of Paraná. Multiplex PCR is an important tool in the detection of Leishmania infection in sandflies.

Cutaneous leishmaniasis (CL) can occur in skin and mucosa, causing disfiguring lesions. The laboratory diagnosis of CL involves immunological methods and optical detection of the parasite, al of which have limitations. There is a need for more effective diagnostic methods for CL which wil allow treatment to be initiated more promptly in order to help prevent the development of severe forms of mucosal disease, and to estimate the prognosis of the infection. The polymerase chain reaction (PCR) has been widely used to diagnose CL, because of its higher sensitivity. This study estimated the accuracy and compared PCRs of samples from lesion scarification (PCR-L) and blood sample-enriched leukocytes (PCR-B) with three conventional diagnostic techniques: parasite direct search (DS), Montenegro skin test (MST), and indirect immunofluorescence reaction (IIF). The study included 276 patients under suspicion of CL. We conducted a cross-sectional study, in which patients were selected by convenience sampling. We used MP3H/MP1L primers to generate a Leishmania (Viannia) (minicircle kDNA) fragment of 70-bp. Of 106 patients with CL, 83.87%, 51.67%, 64.52%, 85.71%, or 96.10% tested positive by PCR-L, PCR-B, DS, IIF, or MST, respectively. Five patients tested positive only by PCR-L, and two other patients only by PCR-B. PCR-L is indicated for use in patients with chronic lesions or Leishmania reinfection, which may progress to mucosal lesion. PCR-B is indicated for use in patients with negative results in conventional tests or for patients with no apparent lesion. PCR is not only useful in diagnosing CL but also helps to identify the infecting species.

Relata-se os resultados de uma investigação sobre a possibilidade da existência de memória em flebotomíneos em área endêmica de leishmaniose tegumentar americana, no Estado do Paraná. Capturaram-se flebotomíneos no Recanto Marista, Doutor Camargo, Estado do Paraná, Brasil, com armadilhas de Falcão, de 15/11/2007 a 26/11/2007, em dois galinheiros (G1 e G2). Foram capturados 2.080 flebotomíneos (1.000 em G1 e 1.080 em G2), que foram marcados e soltos. Nyssomyia neivai foi a espécie mais (90,5%) freqüente. Do total solto, recapturaram-se 168 (8%) flebotomíneos e a recaptura no G2 foi significativa. Os resultados evidenciam que é possível a existência de memória espacial, olfativa e/ou a fidelidade ao hospedeiro e que isto orientem os flebotomíneos no reconhecimento dos locais onde há disponibilidade de fontes de sangue.

RESUMO

Due to the importance of sandflies in the American cutaneous leishmaniasis epidemiology, we investigated the feeding preference of sandflies related to domestic animals common in domiciliary environments in endemic rural areas, where this disease occurs. The sandflies were collected with Falcão light traps, installed in cages, each one containing one domestic animal kind (pig, dog, rabbit or chicken), in Recanto Marista, in Doutor Camargo municipality. Forty five collections were made, totaling 135h of collections per trap. A total of 2,854 sandflies of the following species were collected: Brumptomyia brumpti (Larousse), Evandromyia correalimai (Martins, Coutinho & Luz), Evandromyia cortelezzii (Brèthes), Expapillata firmatoi (Barreto, Martins & Pellegrino), Micropygomyia ferreirana (Barreto, Martins & Pellegrino), Migonemyia migonei (França), Nyssomyia neivai (Pinto), Nyssomyia whitmani (Antunes & Coutinho), Pintomyia fischeri (Pinto), Pintomyia monticola (Costa Lima), Pintomyia pessoai (Coutinho & Barreto), Psathyromyia lanei (Barreto & Coutinho) and Psathyromyia shannoni (Dyar). The dominant species was N. neivai. N. neivai and N. whitmani did not show any feeding preference towards the animals used as bait. It was concluded that N. neivai and N. whitmani are opportunist and, probably, the females adjust their feeding habits to the availability of hosts, suggesting their feeding eclecticism in the anthropic environments.

We report the results of control measures introduced to reduce the density of sand flies in domiciles and subsequent monitoring of the effects of these measures on the sand fly populations. The most common species of sand flies were Nyssomyia neivai and Nyssomyia whitmani, which are naturally infected by Leishmania. A total of 268,382 (93.4%) sand flies were collected in ecotypes constructed with the aim of attracting sand flies, and 19,091 (6.6%) sand flies were collected in the ecotypes consisting of residences and other buildings. Human actions determine the growth or reduction of the sand fly population in human-occupied space. Understanding the dynamics of sand flies in this environment can substantially contribute to the prevention of cutaneous leishmaniasis. Journal of Vector Ecology 38 (1): 63-68. 2013.

We report the results of an investigation of natural larval sand fly habitats in the Recanto Marista, Doutor Camargo municipality, Paraná state, Brazil, from May, 2010 to August, 2012. We used Alencar emergence traps (AT), experimental traps (ET), and soil samples incubated in a biochemical oxygen demand incubator. Eight sand flies were collected with ATs. One specimen was collected with an ET and 21 were collected in soil samples. The collected species were Brumptomyia brumpti, Micropygomyia ferreirana, Migonemyia bursiformis, Migonemyia migonei, Nyssomyia neivai, Nyssomyia whitmani, and Pintomyia pessoai. The laval habitats of sand flies were located in the Recanto Marista, especially between tree roots, but the number of adults that emerged in the traps and soil samples was small despite the high density of sand flies that has been recorded in the Recanto Marista. Journal of Vector Ecology 40 (2): 269-276. 2015.

Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.