Leishmania amazonensis DNA in wild females of Lutzomyia cruzi (Diptera: Psychodidae) in the state of Mato Grosso do Sul, Brazil.

Studies on natural infection by Leishmania spp of sandflies collected in endemic and nonendemic areas can provide important information on the distribution and intensity of the transmission of these parasites. This study sought to investigate the natural infection by Leishmaniain wild female sandflies. The specimens were caught in the city of Corumbá, state of Mato Grosso do Sul (Brazil) between October 2012-March 2014, and dissected to investigate flagellates and/or submitted to molecular analysis to detect Leishmania DNA. A total of 1,164 females (77.56% of which were Lutzomyia cruzi) representing 11 species were investigated using molecular analysis; 126 specimens of Lu. cruziwere dissected and also submitted to molecular analysis. The infection rate based on the presence of Leishmania DNA considering all the sandfly species analysed was 0.69%; only Leishmania (Leishmania) amazonensis was identified in Lu. cruzi by the molecular analysis. The dissections were negative for flagellates. This is the first record of the presence of L. (L.) amazonensis DNA in Lu. cruzi, and the first record of this parasite in this area. These findings point to the need for further investigation into the possible role of this sandfly as vector of this parasite.

Development of Immunological Assays Based on Leishmania donovani Antigen for Diagnosis of Canine Visceral Leishmaniasis and Their Multicenter Evaluation in Brazil and Italy.

Canine visceral leishmaniasis (CVL) due to Leishmania infantum infection is a zoonotic disease prevalent in the areas of South America and the Mediterranean. Infected dogs as reservoirs can contribute to disease transmission and can be a scourge to public health. Therefore, early diagnosis of infected dogs may play a pivotal role in circumscribing disease progression. Invasive tissue aspiration and insufficient serological methods impair a single assay for prompt CVL diagnosis. In the present study, we aimed to evaluate the potential of Leishmania donovani isolated membrane protein, LAg, for the diagnosis of CVL through immunological assays. Initially, enzyme-linked immunosorbent assay was done with Brazilian dog sera to evaluate the performance of LAg in diagnosing CVL and found sensitivity and specificity of 92.50% and 95%, respectively. The study further confirmed the diagnostic efficacy of LAg in a dipstick format. The dipstick test of canine sera from three centers in Brazil and one center in Italy collectively showed sensitivity values in the range of 53.33% to 100% in recognizing symptomatic dogs and specificity values between 75% and 100% to rule out healthy dogs. Moreover, a rapid immunochromatographic test was developed and optimized using LAg. This test was able to identify 94.73% of CVL of Brazilian origin with specificity of 97.29%. The current results highlight the reactive potential of the L. donovani antigen, LAg, for L. infantum CVL diagnosis and support our previous findings, which suggest the utility of LAg for the diagnosis of both L. donovani and L. infantum human VL in a variety of endemic regions. LAg as a diagnostic candidate may be employed to identify comprehensive CVL cases in epidemiological areas.

Canine Visceral Leishmaniasis in an Area of Sporadic Transmission in Brazil.

Canine visceral leishmaniasis is a zoonotic disease caused by the protozoan Leishmania infantum in Latin America. Visceral leishmaniasis (VL) diagnosis in Brazil includes two serological tests according to the Ministry of Health (MH) protocol. Sensitivity and specificity of diagnostic tests, as well as clinical signs of VL, are usually reported in disease-endemic areas; however, it is known that local epidemiological factors can influence these results. This study aimed to evaluate the clinical features, sensitivity, and specificity of TR-DPP® and EIE-LVC in naturally infected dogs in a region of sporadic VL transmission to humans in Brazil. A total of 288 dogs were clinically evaluated and serological and parasitological (lymph node aspirates) samples were collected for VL diagnosis. TR-DPP and EIE-LVC showed poor sensitivity (0.62 and 0.44, respectively) to detect infected animals, compared with the direct parasitological examination, which is considered a gold standard method. Thus, the protocol of MH presented low sensitivity (0.42) to estimate prevalence and control measures in this region. TR-DPP presented a high negative predictive value (0.89), resulting in its indication as a confirmatory test in sporadic transmission areas. Classical clinical signs of VL were not frequently observed; therefore, clinical scoring systems might not be useful in this region. Veterinarians of nonendemic areas should be alert for asymptomatic dogs, especially those presenting lymph adenomegaly.

Giant cutaneous cyst in a dog infected by Cercopithifilaria bainae.

Cercopithifilaria bainae is the most prevalent species of filarioids within the genus. This parasite localizes in the skin, sometimes causing erythematous dermatitis. Herein, the authors describe a case of giant cutaneous cyst in a dog infected by Cercopithifilaria bainae. A 9-year-old male mixed-breed dog presented to a veterinary facility in Dourados, Mato Grosso do Sul (Midwest Brazil) with a mass in the lumbosacral region. On clinical examination, the mass was observed to be approximately 15 cm in diameter with a floating consistency and conspicuous presence of viscous fluid; the lesion, however, was non-ulcerated and non-adherent. Cytological examination revealed the presence of moderate lymphocyte cellularity and foamy macrophages, erythrophagocytosis and the presence of numerous microfilariae. After morphological and molecular analysis of the 12S ribosomal RNA gene, the microfilariae were identified as C. bainae, exhibiting 99-100% identity with DNA sequences available from Genbank. Surgery was recommended and after resection of the giant cyst, the dog was treated with ivermectin for two weeks and the clinical condition was completely resolved. Based on the presence of microfilariae in the cyst fluid the role of this filarioid in the determinism of the lesion has been discussed.

Spatiotemporal analysis of sandfly fauna (Diptera: Psychodidae) in an endemic area of visceral leishmaniasis at Pantanal, central South America.

BACKGROUND: Environmental changes caused by urbanization can cause alterations in the ecology and behavior of sandflies and in the epidemiology of leishmaniasis. Geotechnological tools allow the analysis and recognition of spatiotemporal patterns by monitoring and mapping risk areas of this vector-borne disease. This study aims to describe the sandfly fauna in the municipality of Corumbá and to compare it with the data described in a three-year period from 1984 to 1986 by Galati. A further aim was to analyze the influence of environmental changes on the composition of the fauna. METHODS: Captures were conducted weekly from April 2012 to March 2013, in intra and peridomicile areas with automatic light traps, from 6:00 pm to 6:00 am. The following indices were calculated for both periods analyzed: Standardized Index of Species Abundance (SISA), Shannon's diversity index (H) and Pielou's index (J). The Normalized Difference Vegetation Index (NDVI) was extracted from a remote sensing LANDSAT-5 image. RESULTS: In total, 7,370 specimens (6,169 males and 1,201 females) were collected, distributed among 12 species. Lutzomyia cruzi was the most frequent species (93,79%) and the first in the ranking of standardized species abundance index in both studies. The dominance of the species Lu. cruzi in the neighborhoods of Maria Leite and Centro was demonstrated by the low equitability index. The neighborhood of Cristo Redentor had the greatest diversity of sandflies in the present study and the second greatest in the study performed by Galati et al. (Rev Saúde Pública 31:378-390, 1997). Analyzing the satellite images and the NDVI from 1984 and 2010, the largest amount of dense vegetation was found in the neighborhood of Cristo Redentor. CONCLUSIONS: It was, therefore, possible to show how changes caused due to urbanization have affected the density and distribution of Lu. cruzi and other species over time. Moreover, the data suggest that different populations of sandflies adapt in different ways according to environmental conditions and the adaptation does not necessarily depends on the presence of high vegetation cover.

Phlebotomine sandflies fauna (Diptera: Psychodidae) at rural settlements in the municipality of Cáceres, State of Mato Grosso, Brazil.

INTRODUCTION: Leishmaniasis is expanding across Brazil, including the State of Mato Grosso (MT). The aim of this study was to characterize the phlebotomine sandfly fauna at threerural settlements located in the municipality of Cáceres, MT, from August 2010 to July 2011. METHODS: Sandfly captures were conducted at the forest border and in intra and peridomicile areas with automatic light traps, biweekly, from 5pm to 6am. RESULTS: Of the 630 sandflies collected, 348 were female, and 282 were male. Captured specimens were distributed across 11 genera - Brumptomyia, Evandromyia, Expapillata, Lutzomyia, Martinsmyia, Micropygomyia, Nyssomyia, Pintomyia, Psathyromyia, Psychodopygus, and Sciopemyia - and 28 species. Among these, six species had not been marked yet in MT (Brumptomyia avellari, Br. mangabeirai, Evandromyia aldafalcaoae, Micropygomyia echinatopharynx, Micropygomyia peresi, and Pa. campograndensis). Nyssomyia whitmani was the most abundant species across ecotopes at all settlements. Interestingly, Lutzomyia longipalpis and Lutzomyia cruzi were found in sympatry. CONCLUSIONS: The disorganized occupation of the environment that is happening at the rural settlements of Mata Comprida, Laranjeira I and II could provide opportunities for the domiciliation of wild populations of phlebotomine, including vectors of leishmaniasis. Therefore, more studies are needed to understand the epidemiology of the disease in these areas and its impact on the human population.

Biological aspects of immature stage of Nyssomyia whitmani (Antunes and Coutinho) (Diptera, Psychodidae, Phlebotominae) in laboratory conditions

Nyssomyia whitmani (Antunes and Coutinho, 1939) has been considered as a complex of cryptic species, and some of the populations of this complex plays an important role in the transmission of Leishmania spp. in Brazil. The present study reports the biological aspects concerning the productivity out of eggs and the development time of the descendants of females obtained in Dourados municipality, Mato Grosso do Sul state. The females were captured with modified electric aspirators, fed in hamsters and further individualized in containers for breeding. At the insectary, temperature and relative humidity were maintained on average of 24.5 °C and 67.3%, respectively. From 944 females 3737 eggs were obtained, 748 (20.0%) evolved to the stage of larvae, and 93 (12.4%) of these reached adult stage. The life cycle lasted 80.6 days and the last larval instar was the longest. The use of a higher protein diet revealed a significant improvement in larval development.

Leishmania amazonensis DNA in wild females ofLutzomyia cruzi (Diptera: Psychodidae) in the state of Mato Grosso do Sul, Brazil

Studies on natural infection by Leishmania spp of sandflies collected in endemic and nonendemic areas can provide important information on the distribution and intensity of the transmission of these parasites. This study sought to investigate the natural infection by Leishmaniain wild female sandflies. The specimens were caught in the city of Corumbá, state of Mato Grosso do Sul (Brazil) between October 2012-March 2014, and dissected to investigate flagellates and/or submitted to molecular analysis to detect Leishmania DNA. A total of 1,164 females (77.56% of which were Lutzomyia cruzi) representing 11 species were investigated using molecular analysis; 126 specimens of Lu. cruziwere dissected and also submitted to molecular analysis. The infection rate based on the presence of Leishmania DNA considering all the sandfly species analysed was 0.69%; only Leishmania (Leishmania) amazonensis was identified in Lu. cruzi by the molecular analysis. The dissections were negative for flagellates. This is the first record of the presence of L. (L.) amazonensis DNA in Lu. cruzi, and the first record of this parasite in this area. These findings point to the need for further investigation into the possible role of this sandfly as vector of this parasite.

Phlebotomine sandflies fauna (Diptera: Psychodidae) at rural settlements in the municipality of Cáceres, State of Mato Grosso, Brazil / Fauna flebotomínica (Diptera: Psychodidae) em assentamentos rurais no Município de Cáceres, Estado de Mato Grosso, Brasil

INTRODUCTION: Leishmaniasis is expanding across Brazil, including the State of Mato Grosso (MT). The aim of this study was to characterize the phlebotomine sandfly fauna at threerural settlements located in the municipality of Cáceres, MT, from August 2010 to July 2011. METHODS: Sandfly captures were conducted at the forest border and in intra and peridomicile areas with automatic light traps, biweekly, from 5pm to 6am. RESULTS: Of the 630 sandflies collected, 348 were female, and 282 were male. Captured specimens were distributed across 11 genera - Brumptomyia, Evandromyia, Expapillata, Lutzomyia, Martinsmyia, Micropygomyia, Nyssomyia, Pintomyia, Psathyromyia, Psychodopygus, and Sciopemyia - and 28 species. Among these, six species had not been marked yet in MT (Brumptomyia avellari, Br. mangabeirai, Evandromyia aldafalcaoae, Micropygomyia echinatopharynx, Micropygomyia peresi, and Pa. campograndensis). Nyssomyia whitmani was the most abundant species across ecotopes at all settlements. Interestingly, Lutzomyia longipalpis and Lutzomyia cruzi were found in sympatry. CONCLUSIONS: The disorganized occupation of the environment that is happening at the rural settlements of Mata Comprida, Laranjeira I and II could provide opportunities for the domiciliation of wild populations of phlebotomine, including vectors of leishmaniasis. Therefore, more studies are needed to understand the epidemiology of the disease in these areas and its impact on the human population.

INTRODUÇÃO: As leishmanioses encontram-se em expansão no Brasil, inclusive no Estado de Mato Grosso (MT). A presente pesquisa teve como objetivo estudar a fauna flebotomínica emtrês assentamentos rurais localizados no município de Cáceres, MT, entre agosto de 2010 e julhode 2011. MÉTODOS: As coletas foram realizadas na margem da mata, no intra e peridomicílio com armadilhas automáticas luminosas, quinzenalmente, das 17h às 6h. RESULTADOS: Foram coletados 630 exemplares, sendo 348 fêmeas e 282 machos. Os espécimes foramdistribuídos em 11 gêneros: Brumptomyia, Evandromyia, Expapillata, Lutzomyia, Martinsmyia,Micropygomyia, Nyssomyia, Pintomyia, Psathyromyia, Psychodopygus e Sciopemyia e 28 espécies, sendo que dentre estas, seis não haviam sido assinaladas em MT (Brumptomyia avellari, Br. mangabeirai, Evandromyia aldafalcaoae, Micropygomyia echinatopharynx, Micropygomyia peresi e Pa. campograndensis). Ressalta-se o encontro de Nyssomyia whitmani, espécie mais abundante nos diferentes ecótopos em todos os assentamentos. Destaca-se também a presençadas espécies Lutzomyia longipalpis e Lutzomyia cruzi em simpatria. CONCLUSÕES: A ocupação desordenada do ambiente, como vem ocorrendo nos assentamentos Mata Comprida, LaranjeiraI e II, pode proporcionar a domiciliação de populações selváticas de flebotomíneos, dentre estes os vetores das leishmanioses, tornando-se necessários maiores estudos para entender a dinâmica de transmissão dessas morbidades nestes locais.