Species-specific identification of Leishmania in naturally infected sand flies captured in Mato Grosso do Sul State, Brazil

An increase in cutaneous and visceral leishmaniasis cases has been reported in recent years in the state of Mato Grosso do Sul, Brazil, and little is known to date about their etiological agents. An investigation into natural Leishmania infection of sand flies captured in this state between December 2003 and August 2004 was carried out. Mini-exon sequences were used as targets to identify Leishmania, and an RFLP technique was employed for those identified as belonging to the Viannia subgenus. Calculation of the minimal infection rate (MR) revealed that 1.6% of sand flies captured in the forest, peridomicile and intradomicile were positive. Six species were found to be infected by Leishmania (V.) braziliensis. Interestingly, two of the six species. Lutzomyia longipalpis and Nyssomyia whitmani, were captured in anthropic environments. The findings of this study constitute a useful tool for planning control measures against this disease in the State of Mato Grosso do Sul. (C) 2010 Elsevier B.V. All rights reserved.

Phylogenetic Analyses Based on Small Subunit rRNA and Glycosomal Glyceraldehyde-3-Phosphate Dehydrogenase Genes and Ultrastructural Characterization of Two Snake Trypanosomes: Trypanosoma serpentis n. sp from Pseudoboa nigra and Trypanosoma cascavelli from Crotalus durissus terrificus

We sequenced the small subunit (SSU) rRNA and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes of two trypanosomes isolated from the Brazilian snakes Pseudoboa nigra and Crotalus durissus terrificus. Trypanosomes were cultured and their morphometrical and ultrastructural features were characterized by light microscopy and scanning and transmission electron microscopy. Phylogenetic trees inferred using independent or combined SSU rRNA and gGAPDH data sets always clustered the snake trypanosomes together in a clade closest to lizard trypanosomes, forming a strongly supported monophyletic assemblage (i.e. lizard-snake clade). The positioning in the phylogenetic trees and the barcoding based on the variable V7-V8 region of the SSU rRNA, which showed high sequence divergences, allowed us to classify the isolates from distinct snake species as separate species. The isolate from P. nigra is described as a new species, Trypanosoma serpentis n. sp., whereas the isolate from C. d. terrificus is redescribed here as Trypanosoma cascavelli.

Spatial and temporal patterns of occurrence of Lutzomyia sand fly species in an endemic area for cutaneous leishmaniasis in the Atlantic Forest region of northeast Brazil

Sand fly populations of different ecological niches in the Amaraji endemic American Cutaneous Leishmaniasis (ACL) focus of the Pernambuco Atlantic Forest region of northeastern Brazil were monitored spatiotemporally. Lutzomyia whitmani was dominant in all niches but occurred in smaller numbers in forested locations. L. whitmani was significantly less seasonal than the other species, being present throughout the year while other species were more abundant between February and April. These results suggest that L. whitmani may potentially be the principal vector of ACL in the region, even though the sand fly fauna was diverse: 88% were L.whitmani and 12% belonged to 11 other species. Two other species, L. complexa (1.3%) and L. migonei (0.8%), considered to be ACL vectors in other regions, were also present. This detailed picture of the sand fly population`s abundance and spatiotemporal distribution provides a basis for future modeling studies of forecasting sand fly activity patterns and ACL occurence.

Phylogeny of snake trypanosomes inferred by SSU rDNA sequences, their possible transmission by phlebotomines, and taxonomic appraisal by molecular, cross-infection and morphological analysis

Blood examination by microhaematocrit and haemoculture of 459 snakes belonging to 37 species revealed 24% trypanosome prevalence in species of Viperidae (Crotalus durissus and Bothrops jararaca) and Colubridae (Pseudoboa nigra). Trypanosome cultures from C. durissus and P. nigra were behaviourally and morphologically indistinguishable. In addition, the growth and morphological features of a trypanosome from the sand fly Viannaniyia tuberculata were similar to those of snake isolates. Cross-infection experiments revealed a lack of host restriction, as snakes of 3 species were infected with the trypanosome from C. durissus. Phylogeny based on ribosomal sequences revealed that snake trypanosomes clustered together with the sand fly trypanosome, forming a new phylogenetic lineage within Trypanosoma closest to a clade of lizard trypanosomes transmitted by sand flies dagger. The clade of trypanosomes from snakes and lizards suggests an association between the evolutionary histories of these trypanosomes and their squamate hosts. Moreover, data strongly indicated that these trypanosomes are transmitted by sand flies. The flaws of the current taxonomy of snake trypanosomes are discussed, and the need for molecular parameters to be adopted is emphasized. To our knowledge, this is the first molecular phylogenetic study of snake trypanosomes.

Biologia alimentar da taxocenose de peixes do Rio Silveira (cabeceira do Rio Pelotas), São José dos Ausentes, Rio Grande do Sul, Brasil

A região de São José dos Ausentes (planalto sul-riograndense), onde se situa o rio Silveira pertencente a cabeceira do rio Pelotas (bacia do rio Uruguai), caracteriza-se por uma ictiofauna composta de espécies endêmicas e várias espécies novas ainda não descritas. O conhecimento referente a estrutura trófica da ictiofauna desta região ainda é inexistente, motivo pelo qual este trabalho teve com objetivo caracterizar a biologia alimentar das espécies de peixes encontradas no rio Silveira. O período de amostragem iniciou em julho de 2000 e concluiu-se em julho 2001. Os indivíduos foram capturados mensalmente através de redes de espera (malhas 1,5 cm entrenós) e puçás, em ambientes de corredeiras e remansos. Foram encontradas seis espécies de peixes neste trecho do rio Silveira: Astyanax sp. n. 4 (Characidae), Eurycheilichthys pantherinus (Loricariidae), Hemipsilichthys sp. n. I (Loricariidae), Cichlasoma facetum (Cichlidae), Rhamdia sp. (Pimelodidae) e Bryconamericus sp. n. 1 (Characidae). O conteúdo estomacal foi identificado para cada uma destas espécies, sendo somente analisado através do método de freqüência de ocorrência, composição percentual e pelo cálculo do índice de importância alimentar para as três espécies mais freqüentes e abundantes. Dentre os itens encontrados destacam-se para Astyanax sp. n. 4 matéria vegetal e Ephemeroptera; para Eurycheilichthys pantherinus larvas de Diptera Simulidae e Ephemeroptera; e em Hemipsilichthys sp. n. I detritos, onde se verificou a presença de diatomáceas e clorofítas. O cálculo do coeficiente de sobreposição alimentar não apresentou valor significativo para estas três espécies. As demais espécies apresentaram os seguintes itens em sua dieta, Cichlasoma facetum larvas de Tricoptera, Ephemeroptera, Gastropoda e insetos alóctones; Rhamdia sp. escamas, Gastropoda, larvas de Lepidoptera, Tricoptera, Ephemeroptera, sementes, matéria vegetal, sedimento; e Bryconamericus sp. n. 1 larvas de Diptera (Simulidae, Psychodidae, Chironomidae), larvas de Tricoptera, Ephemeroptera, Diatomáceas, Clorofítas, Diptera adulto, Hymenoptera, Coleoptera. Esta dieta sugere os seguintes hábitos alimentares para Astyanax sp. n. 4 onívoro, Eurycheilichthys pantherinus insetívoro, Hemipsilichthys sp. n. I detritívoro, Cichlasoma facetum insetívoro, Rhamdia sp. onívoro, e Bryconamericus sp. n. 1 insetívoro.

Genetic Structure of Lutzomyia longipalpis Populations in Mato Grosso Do Sul, Brazil, Based on Microsatellite Markers

Background:Lutzomyia longipalpis (Diptera: Psychodidae) is the major vector of Leishmania (Leishmania) infantum and thus plays a crucial role in the epidemiology of American visceral leishmaniasis (AVL). This vector is the best studied species of sand fly in the Neotropical region. Many studies claim that this vector is in fact a species complex; however there is still no consensus regarding the number of species that belong into this complex or the geographical distribution of sibling species. The aim of the present study was to analyze the genetic relationships within Lu. longipalpis populations in the state of Mato Grosso do Sul (MS), Brazil.Methodology/Principal Findings:We collected 30 Lu. longipalpis (15 females and 15 males) from five localities (Campo Grande, Três Lagoas, Aquidauana, Miranda and Bonito) and 30 Lu. Cruzi from Corumbá, totaling 180 sandflies from MS, and 30 Lu. longipalpis from Estrela de Alagoas, state of Alagoas (AL), Northeast Brazil. We show that eight previously described microsatellite loci were sufficient in distinguishing Lu. longipalpis from Lu. Cruzi, which is a closely related species, and in differentiating between Lu. longipalpis collected in MS versus Estrela de Alagoas. Analyses of the genotypes revealed introgression between sympatric Lu. longipalpis and Lu. Cruzi.Conclusions/Significance:Our findings support the hypothesis of cryptic species within the Lu. longipalpis complex. Furthermore, our data revealed introgression between Lu. longipalpis and Lu. cruzi. This phenomenon should be further investigated to determine the level and incidence of hybridization between these two species. We also demonstrated that microsatellite markers are a powerful tool for differentiating sand fly populations and species. The present study has elucidated the population structure of Lu. longipalpis in MS and, by extension, the Neotropical Lu. longipalpis complex itself. © 2013 Santos et al.

Avaliação da diversidade de insetos hematófagos da subordem Nematocera e de vertebrados silvestres: transmissão de arbovírus na área de influência do Projeto Salobo, Carajás, Pará

Com o objetivo de avaliar a diversidade de insetos hematófagos e de vertebrados silvestres, bem como, a fauna de arbovírus circulante antes das ações de exploração mineral na jazida polimetálica do Salobo, Província Mineral de Carajás, Pará, Brasil, no período de dezembro de 2005 a junho de 2007, um estudo longitudinal foi realizado (sete viagens) sendo capturados e identificados insetos hematófagos (famílias Ceratopogonidae, Culicidae, Psychodidae e Simulidae) capturados em armadilhas luminosas CDC e Shannon, e atração humana; e também foram capturados e identificados vertebrados silvestres das classes das aves (redes de nylon), dos mamíferos e dos répteis (armadilhas Shermann e Tommahwak); foi feita pesquisa e determinação da prevalência de anticorpos nos soros e/ou plasmas desses vertebrados contra arbovírus e tentativas de isolamento viral. Foram capturados 44.795 (1.220 lotes) insetos hematófagos, sendo a família Psychodidae a mais prevalente. As espécies mais abundantes de culicídeos foram Haemagogus leucocelaenus e Haemagogus janthinomys. Foram também capturados 1.288 vertebrados silvestres, e os roedores Proechimys guyannensis e Oryzomys capito, e as aves Turdus albicollis e Phlegopsis nigromaculata foram as espécies mais prevalentes. Foram isoladas em camundongos recém-nascidos, três cepas do Virus Tucunduba, obtidas a partir de lotes de Anopheles (Nys.) species, Culex coronator e Wyeomyia species; foram detectados anticorpos para os seguintes arbovírus: encefalite Saint Louis (VSLE), Ilhéus, encefalite eqüina Oeste, Cacipacoré, Icoaraci, Rocio, Bussuquara e Mucambo, sendo a maior prevalência de anticorpos obtida para o VSLE.

Utilização de armadilha de emersão para captura de flebotomíneos em área com transmissão e sem transmissão de LVA e LTA, na região centro-oeste do estado de São Paulo, Brasil

Pós-graduação em Medicina Veterinária - FMVZ

Fatores de riscos associados à leishmaniose visceral canina na área de cinturão verde de Ilha Solteira, SP

Pós-graduação em Ciência e Tecnologia Animal - FEIS

Leishmania (Viannia) braziliensis is the main species causing cutaneous leishmaniasis in the Federal District of Brazil

The first autochthonous case of American cutaneous leishmaniasis was reported in the Federal District in 1980, and the species involved in this type of leishmaniasis was unknown. This study aimed to identify the species that causes the disease in the Federal District and to investigate its clinical and epidemiological aspects. Between 2000 and 2007, 71 autochthonous cases of leishmaniasis were reported in the Federal District. Leishmania species were identified by means of direct immunofluorescence reactions using monoclonal antibodies and restriction fragment length polymorphism. The species of 40 (56.33%) out of 71 samples were identified. Thirty-six (90%) were identified as Leishmania (Viannia) braziliensis and four (10%) were identified as Leishmania (Leishmania) amazonensis. In this area, the disease had clinical and epidemiological characteristics similar to those found in other Brazilian regions.

Space-time cluster analysis of American visceral leishmaniasis in Bauru, Sao Paulo State, Brazil

A space-time analysis of American visceral leishmaniasis (AVL) in humans in the city of Bauru, Sao Paulo State, Brazil was carried out based on 239 cases diagnosed between June 2003 and October 2008. Spatial analysis of the disease showed that cases occurred especially in the city's urban areas. AVL annual incidence rates were calculated, demonstrating that the highest rate occurred in 2006 (19.55/100,000 inhabitants). This finding was confirmed by the time series analysis, which also showed a positive tendency over the period analyzed. The present study allows us to conclude that the disease was clustered in the Southwest side of the city in 2006, suggesting that this area may require special attention with regard to control and prevention measures.

Macroinvertebrate abundance and proportion of trait categories in river biotopes across England and Wales

There is a long tradition of river monitoring using macroinvertebrate communities to assess environmental quality in Europe. A promising alternative is the use of species life-history traits. Both methods, however, have relied on the time-consuming identification of taxa. River biotopes, 1-100 m**2 'habitats' with associated species assemblages, have long been seen as a useful and meaningful way of linking the ecology of macroinvertebrates and river hydro-morphology and can be used to assess hydro-morphological degradation in rivers. Taxonomic differences, however, between different rivers had prevented a general test of this concept until now. The species trait approach may overcome this obstacle across broad geographical areas, using biotopes as the hydro-morphological units which have characteristic species trait assemblages. We collected macroinvertebrate data from 512 discrete patches, comprising 13 river biotopes, from seven rivers in England and Wales. The aim was to test whether river biotopes were better predictors of macroinvertebrate trait profiles than taxonomic composition (genera, families, orders) in rivers, independently of the phylogenetic effects and catchment scale characteristics (i.e. hydrology, geography and land cover). We also tested whether species richness and diversity were better related to biotopes than to rivers. River biotopes explained 40% of the variance in macroinvertebrate trait profiles across the rivers, largely independently of catchment characteristics. There was a strong phylogenetic signature, however. River biotopes were about 50% better at predicting macroinvertebrate trait profiles than taxonomic composition across rivers, no matter which taxonomic resolution was used. River biotopes were better than river identity at explaining the variability in taxonomic richness and diversity (40% and <=10%, respectively). Detailed trait-biotope associations agreed with independent a priori predictions relating trait categories to near river bed flows. Hence, species traits provided a much needed mechanistic understanding and predictive ability across a broad geographical area. We show that integration of the multiple biological trait approach with river biotopes at the interface between ecology and hydro-morphology provides a wealth of new information and potential applications for river science and management.

Leishmaniose visceral americana: avaliação dos parâmetros da capacidade vetorial de Lutzomyia longipalpis em área urbana do muncípio de Panorama, São Paulo, Brasil

Introdução: A leishmaniose visceral (LV) é um importante problema de saúde pública no Brasil, com cerca 3000 mil casos notificados anualmente. Nos últimos anos, a LV tem ampliado sua distribuição em vários estados do país, associada principalmente aos processos socioambientais, antrópicos e migratórios. A LV é causada pela infecção com Leishmania infantum chagasi, transmitida, principalmente, por Lutzomyia longipalpis (Diptera: Psychodidae). Este flebotomíneo apresenta ampla distribuição nas Américas, todavia, evidências sugerem que se constitui em um complexo de espécies crípticas. A dinâmica de transmissão da LV é modulada por fatores ecológicos locais que influenciam a interação entre populações do patógeno, do vetor e dos hospedeiros vertebrados. Portanto, o estudo das variáveis associadas a esta interação pode contribuir para elucidar aspectos dos elos epidemiológicos e contribuir para a tomada de decisões em saúde pública. Objetivo: Avaliar parâmetros relacionados à capacidade vetorial da população de Lu. longipalpis presente em área urbana do município de Panorama, estado de São Paulo. Métodos: Foram realizadas capturas mensais durante 48 meses para avaliar a distribuição espaço-temporal de Lu. longipalpis e investigar a circulação de Le. i. chagasi. Também foram realizados os seguintes experimentos com o vetor: captura-marcação-soltura-recaptura para estimar a sobrevida da população e a duração do seu ciclo gonotrófico, a atratividade dos hospedeiros mais frequentes em áreas urbanas, a proporção de repasto em cão, infecção experimental e competência vetorial. Resultados: Observou-se que no município de Panorama, Lu. longipalpis apresentou as frequências mais elevadas na estação chuvosa (entre outubro e março), maior densidade em áreas com presença de vegetação e criação de animais domésticos, locais aonde também foi demonstrada a circulação natural de espécimes de Lu. longipalpis infectados com Le. i. chagasi. Além disto, foi corroborado que a população de Lu. longipalpis apresentou hábito hematofágico eclético, altas taxas de sobrevivência e que foi competente para transmitir o agente da LV. Nos experimentos de laboratório foi evidenciada a heterogeneidade na infecção de fêmeas de Lu. longipalpis desafiadas a se alimentarem em cães comprovadamente infectados por L. i. chagasi e o rápido desenvolvimento do parasita neste vetor natural. Conclusões. As observações do presente estudo corroboram a capacidade vetora de Lu. longipalpis para transmitir a Le. i. chagasi e ressaltam a importância da espécie na transmissão do agente etiológico da LV. Ações de manejo ambiental, educação e promoção à saúde são recomendadas às autoridades municipais para diminuir o risco potencial de infecção na população humana e canina, considerando-se o elevado potencial vetor de Lu. longipalpis e a presença de condições que favorecem a interação dos componentes da tríade epidemiológica da LV.

Colombia, el país con más especies de parásitos de Leishmania

Un reciente estudio pone en evidencia que el país tiene nueve especies de parásitos que producen la leishmaniasis, eso lo convierte en la nación que más tiene en todo el mundo. Investigadores piden capacitar a las Secretarías de Salud donde la dolencia es endémica para que aprendan a identificarlas.