Morphological, morphometric, and molecular characterization of Hepatozoon spp. (Apicomplexa, Hepatozoidae) from naturally infected Caudisona durissa terrifica (Serpentes, Viperidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Population structure of the malaria vector Anopheles darlingi in Rondônia, Brazilian Amazon, based on mitochondrial DNA

Anopheles darlingi is the most important Brazilian malaria vector, with a widespread distribution in the Amazon forest. Effective strategies for vector control could be better developed through knowledge of its genetic structure and gene flow among populations, to assess the vector diversity and competence in transmitting Plasmodium. The aim of this study was to assess the genetic diversity of An. darlingi collected at four locations in Porto Velho, by sequencing a fragment of the ND4 mitochondrial gene. From 218 individual mosquitoes, we obtained 20 different haplotypes with a diversity index of 0.756, equivalent to that found in other neotropical anophelines. The analysis did not demonstrate significant population structure. However, haplotype diversity within some populations seems to be over-represented, suggesting the presence of sub-populations, but the presence of highly represented haplotypes complicates this analysis. There was no clear correlation among genetic and geographical distance and there were differences in relation to seasonality, which is important for malarial epidemiology.

Population dynamics, structure and behavior of Anopheles darlingi in a rural settlement in the Amazon rainforest of Acre, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The opportunistic feeding and reproduction strategies of the annual fish Cynopoecilus melanotaenia (Cyprinodontiformes: Rivulidae) inhabiting ephemeral habitats on southern Brazil

A maioria dos peixes da família Rivulidae são popularmente conhecidos como anuais por completarem todo seu ciclo biológico em pequenos corpos de água temporários que secam obrigatoriamente em determinados períodos do ano causando a morte dos indivíduos adultos. Possuem características biológicas peculiares como pequeno porte, maturação sexual precoce, reprodução contínua, um elaborado padrão de corte e uma grande capacidade reprodutiva entre os peixes. Os rivulídeos se encontram amplamente distribuídos nas Américas do Norte, Central e Sul. Este trabalho analisou a dieta e a biologia reprodutiva de Cynopoecilus melanotaenia. Foram coletados 263 exemplares e a análise de 233 conteúdos gastrointestinais revelou uma dieta invertívora composta principalmente por microcrustáceos (Cladocera, Amphipoda e Ostracoda) e insetos imaturos (Chaoboridae, Culicidae, Syrphidae, mas principalmente larvas de Chironomidae). Foi registrada a lepidofagia na dieta dos machos. A fecundidade foi estimada pela análise de 59 pares de ovários maduros e variou entre 2 a 157 ovócitos (média, 19 ± 26[DP]). A espécie apresentou desova parcelada, uma estratégia para aumentar a chance de sobrevivência a depleções prolongadas. Este trabalho apresenta a primeira investigação sobre a biologia reprodutiva de C. melanotaenia. Os resultados encontrados sugerem a estratégia reprodutiva oportunista de C. melanotaenia e fornecem novas informações biológicas que podem contribuir para pesquisas de conservação e manutenção da espécie em seu ambiente natural.

Optimal and sub-optimal control in Dengue epidemics

This work concerns the application of the optimal control theory to Dengue epidemics. The dynamics of this insect-borne disease is modelled as a set of non-linear ordinary differential equations including the effect of educational campaigns organized to motivate the population to break the reproduction cycle of the mosquitoes by avoiding the accumulation of still water in open-air recipients. The cost functional is such that it reflects a compromise between actual financial spending (in insecticides and educational campaigns) and the population health (which can be objectively measured in terms of, for instance, treatment costs and loss of productivity). The optimal control problem is solved numerically using a multiple shooting method. However, the optimal control policy is difficult to implement by the health authorities because it is not practical to adjust the investment rate continuously in time. Therefore, a suboptimal control policy is computed assuming, as the admissible set, only those controls which are piecewise constant. The performance achieved by the optimal control and the sub-optimal control policies are compared with the cases of control using only insecticides when Breteau Index is greater or equal to 5 and the case of no-control. The results show that the sub-optimal policy yields a substantial reduction in the cost, in terms of the proposed functional, and is only slightly inferior to the optimal control policy. Copyright (C) 2001 John Wiley & Sons, Ltd.

Dirofilariose: zoonose emergente negligenciada

A dirofilariose é uma zoonose pouco conhecida causada por Dirofilaria spp., nematódeo mais conhecido como verme do coração dos cães (Dirofilaria immitis), parasita do sistema circulatório desses animais, mas que também pode acometer gatos e o ser humano. Sua ocorrência está intimamente ligada à presença de mosquitos vetores (Aedes spp., Anopheles spp., Culex spp.), condições climáticas favoráveis, assim como trânsito entre regiões indenes e endêmicas/epidêmicas. O ser humano pode se infectar com D. immitis (pulmão), Dirofilaria repens (pulmão, subcutâneo) e Dirofilaria tenuis (subcutâneo). A fisiopatologia está intimamente ligada à morte do parasita onde, no cão, pode induzir a obstrução de vasos circulatórios e no ser humano produzir uma lesão nodular com intensa reação inflamatória no parênquima pulmonar com formato de moeda observada nas radiografias. Pode ser diagnosticada pelo exame físico, pela detecção de microfilárias na circulação sangüínea, imunoadsorção enzimático (ELISA), alterações radiográficas, ecocardiografia, ultrassonografia e necropsia. Há riscos no tratamento, sendo a prevenção com a utilização de drogas nos animais o método mais eficaz, principalmente em visitas a áreas endêmicas ou epidêmicas, diminuindo-se, assim, o risco para saúde pública devido à disseminação do parasita.

Os efeitos da ação antrópica sobre a distribuição de macroinvertebrados no Rio Guareí, São Paulo

In this study, it was examined, during the period from March to December 2006, the effects of human disturbance on themacroinvertebrates that live near macrophytes in Guare River, São Paulo State - Brazil. It was questioned if the high conductivityrecorded in Guare River affected the distribution of the macroinvertebrates and what were the most important variables thataffect macroinvertebrates in a river with a strong nutrient concentration. The objective of this study was to investigate theeffects of environmental variables on densities and composition of the macroinvertebrates. Three stands of aquatic plantswere sampled with with 0.25 mm mesh net on a 0.07 m2 square metal frame. Air and water temperature, depth, pH, electricalconductivity, suspended solids, dissolved oxygen and macrophyte biomass were measured. A canonical correspondence analysis(CCA) was performed using the density of the macroinvertebrates and environmental variables. Chironomidae, Culicidae,Acanthagrion, Coryphaeschna, Erythrodiplax, Miathyria marcella, Micrathyria, Gastropoda, Ostracoda and Hemiptera werethe only taxa that showed significant correlation with the axes. From the results, we can conclude that the high conductivity recordedin Guare River due to the high amount of organic matter released during its course did not significantly affected thedistribution of the macroinvertebrates during the studied period. However, the ACC recorded that oxygen was the most significantenvironmental factor for the density variance of the macroinvertebrates, especially larval Odonata.

Heterochromatic banding pattern in two Brazilian populations of Aedes aegypti

We analysed samples of Aedes aegypti from Sao Jose do Rio Preto and Franca (Brazil) by C-banding and Ag-banding staining techniques. C-banding pattern of Ae. aegypti from Sao Jose do Rio Preto examined in metaphase cells differed from Franca. The chromosomes 2, 3 and X showed centromeric C-bands in both populations, but a slightly stained centromeric band in the Y chromosome was observed only in Sao Jose do Rio Preto. In addition, the X chromosome in both populations and the Y chromosome of all individuals from Sao Jose do Rio Preto showed an intercalary band on one of the arms that was absent in Franca. An intercalary, new band, lying on the secondary constriction of chromosome 3 was also present in mosquitoes of both populations. The comparison of the present data with data in the literature for Ae. aegypti from other regions of the world showed that they differ as to the banding pattern of sex chromosomes and the now described intercalary band in chromosome 3. The observations suggested that the heterochromatic regions of all chromosomes are associated to constitute a single C-banded body in interphase cells. Ag-banding technique stained the centromeric regions of all chromosomes (including the Y) and the intercalary C-band region of the X chromosome in both populations. As Ae. aegypti populations are widespread in a great part of the world, the banding pattern variations indicate environmental interactions and may reveal both the chromosome evolutionary patterns in this species and the variations that may interfere with its vector activity.

Morphological aspects of Culex quinquefasciatus salivary glands

The salivary glands of Culex quinquefasciatus female mosquitoes are paired organs composed of two lateral lobes with proximal and distal secretory portions, and a medial lobe. All portions comprise a simple epithelium that surrounds a salivary duct. In the apical portion of the medial lobe, non-secretory cells strongly resemble cells involved in ion and water transport. The general architecture of the secretory portions is similar between lobes. The appearance of the secretory material and the morphological aspect of the apical cell membrane are the most distinctive features among the three secretory portions. Cells in the lateral proximal lobe display thin membrane projections extending into a translucent and finely filamentous secretory product. At the lateral distal portion, the apical cell membrane forms an intricate meshwork that encloses a dark secretory product. Medial lobe secretory cells also contain secretory cavities surrounded by intracytoplasmic vesicles, all containing a very dark and uniform product. Scattered cells holding numerous vacuoles, some of them containing a small and electron-dense granule eccentrically located and resembling those of the diffuse endocrine system, are frequently observed in the periphery of all secretory portions. Immunofluorescence assays revealed that the distal portion of the lateral lobes contains apyrase, an enzyme putatively responsible for platelet aggregation inhibition, diffusely distributed in the cell cytoplasm. © 2003 Elsevier Ltd. All rights reserved.

Two Digestive Trypsins Occur in Three Species of Neotropical Anophelines

Trypsin activity increases in the midgut of Anopheles aquasalis, Anopheles albitarsis, and Anopheles darlingi after a bloodmeal. The activity returns to basal levels at the time the blood is completely digested. Affinity chromatography, reversed-phase high performance liquid chromatography (HPLC), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to sequentially purify the mosquito trypsins found in the midguts at 24 h after feeding. Amino-terminal sequencing of the purified trypsins showed the occurrence of two distinct trypsins in the midgut of each of the mosquitoes studied. The sequences obtained are similar to those of the trypsins of other hematophagous insects.

Efficacy of topical permethrin as repellent against aedes aegypti's bites

Mosquitoes are the most important vectors of infectious diseases and their bites are related to several adverse skin reactions. Permethrin impregnated clothes are an efficient strategy against arthropods' bites; however, its topical efficacy as a repellent has not been well established. We studied the response to permethrin lotion 5 percent and N,N-Diethyl-meta-toluamide (DEET) spray 50 percent applied to the unprotected forearms of 10 volunteers. Each arm was exposed to 20 female mosquitoes of Aedes aegypti. We performed 71 bilateral comparative measurements evaluating the timing for the first bites. The average times for the arm without the product, with permethrin 5 percent, and with DEET 50 percent were: 7.9 seconds, 336.2 seconds and 7512.1 seconds. The results showed a significant difference between repellency times between either product and unprotected controls. In addition, there was a significant difference in time to first bite between permethrin and DEET treated arms (p<0.01). Permethrin affords some repellent activity against Aedes aegypti bites in this experimental setting. However, permethrin's profile of repellency was significantly inferior to that of DEET. © 2008 Dermatology Online Journal.

Macroinvertebrados aquáticos associados à Eichhornia azurea (Swartz) Kunth e suas relações com as variáveis abióticas em ecossistemas lênticos marginais (São Paulo, Brasil)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The Genome of Anopheles darlingi, the main neotropical malaria vector

Anopheles darlingi is the principal neotropical malaria vector, responsible for more than a million cases of malaria per year on the American continent. Anopheles darlingi diverged from the African and Asian malaria vectors ∼100 million years ago (mya) and successfully adapted to the New World environment. Here we present an annotated reference A. darlingi genome, sequenced from a wild population of males and females collected in the Brazilian Amazon. A total of 10 481 predicted protein-coding genes were annotated, 72% of which have their closest counterpart in Anopheles gambiae and 21% have highest similarity with other mosquito species. In spite of a long period of divergent evolution, conserved gene synteny was observed between A. darlingi and A. gambiae. More than 10 million single nucleotide polymorphisms and short indels with potential use as genetic markers were identified. Transposable elements correspond to 2.3% of the A. darlingi genome. Genes associated with hematophagy, immunity and insecticide resistance, directly involved in vectorhuman and vectorparasite interactions, were identified and discussed. This study represents the first effort to sequence the genome of a neotropical malaria vector, and opens a new window through which we can contemplate the evolutionary history of anopheline mosquitoes. It also provides valuable information that may lead to novel strategies to reduce malaria transmission on the South American continent. The A. darlingi genome is accessible at www.labinfo.lncc.br/index.php/anopheles- darlingi. © 2013 The Author(s).

Genetic structure of Aedes aegypti populations determined using pairwise comparisons

The biological characteristics of Aedes aegypti (Diptera, Culicidae), which is a vector of dengue and yellow fever, make this organism a good model for studying population structure and the events that may influence it under the effect of human activity. We assessed the genetic variability of five A. aegypti populations using RAPD-PCR technique and six primers. Four populations were from Brazil and one was from the USA. A total of 165 polymorphic DNA loci were generated. Considering the six primers and the five populations, the mean value of inter-population genetic diversity (Gst) was 0.277, which is considered high according to the Wright classification. However, pairwise comparisons of the populations gave variable Gst values ranging from 0.044 to 0.289. This variation followed the population's geographic distance to some extent but was also influenced by human activity. The lowest Gst values were obtained in the comparison of populations from cities with intensive commercial and medical contacts. These mosquito populations were previously classified as insecticide resistant, susceptible, or with decreased susceptibility; this parameter apparently had an effect on the Gst values obtained in the pairwise comparisons. ©FUNPEC-RP.

CD95 (FAS) e CD178 (FASL) induzem apoptose em células CD4+ E CD8+ do sangue periférico e esplênicas em cães naturalmente infectados por Leishmania spp: Kathlenn Liezbeth Oliveira Silva. -

Pós-graduação em Ciência Animal - FMVA