Análise da expressão e silenciamento de genes do trato digestivo de Anopheles aquasalis

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

Predição in vitro da atividade tóxica de isolados de Bacillus thuringiensis Berliner e efeito sinergístico no controle de larvas de Aedes aegypti (L.) (Diptera: Culicidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação de pepineiro enxertado em diferentes ambientes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação in vitro dos possíveis efeitos citotóxicos e genotóxicos do alcalóide julocrotina em linfócitos humanos

A leishmaniose é considerada um problema de saúde pública, principalmente devido à presença de diferentes espécies enzoóticas de Leishmania, envolvendo muitos hospedeiros e diferentes insetos vetores. Os antimoniais pentavalentes permanecem como as drogas de primeira escolha para o tratamento das leishmanioses há mais de 50 anos, no entanto, sua utilização vem sendo comprometida devido, principalmente, a resistência desenvolvida pelo parasita ao medicamento. Assim, a escolha de drogas derivadas de plantas baseada no estudo e utilização de práticas da medicina tradicional devem aparecer como nova estratégia para o controle da leishmaniose. No entanto, é importante verificar que algumas destas drogas podem ser tóxicas ao organismo, podendo inclusive apresentar propriedades genotóxicas, causando alterações no DNA com conseqüente aumento no risco de carcinogênese. A Julocrotina (2-[N-(2-methylbutanolyl)]- N-phenylethylglutarimide) é um alcalóide glutarimida isolado da espécie Croton pullei var. glabrior Lanj. Euphorbiaceae), encontrada amplamente na Floresta Amazônica e conhecido por possuir potente efeito leishmanicida. Desta forma, no presente estudo avaliamos o efeito citotóxico e genotóxico da Julocrotinaa partir do Ensaio do MTT e Ensaio do Cometa (versão alcalina) em cultura de linfócitos humanos. Como resultado, o alcalóide não demonstrou citotoxicidade em linfócitos humanos nas concentrações testadas. No entanto, a julocrotina mostrou-se genotóxica para linfócitos humanos tratados com a maior concentração (632μM) da substância. Apesar dos resultados de citotoxicidade parecem promissores no que diz respeito ao uso da julocrotina no tratamento da leishmaniose, o efeito genotóxico observado reforça a necessidade de se obter as devidas precauções quanto ao seu uso como fitoterápico leishmanicida.

Coloration of the testicular peritoneal sheath as a synapomorphy of triatomines (Hemiptera, Reduviidae)

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

Leishmaniose visceral: situação atual e controle

In Brazil, the importance of leishmaniosis it is seen in its high incidence and huge distribution, as well as the possibility of became a lethal disease. It´s a zoonosis of canids, felines, rodents e marsupials, that is transmitted to men through vector bugs. Visceral leishmaniosis, the most serious and prevalent form, strongly became as an emergent and endemic disease, seeing in many Brazilian states, also bringing a lot of problems to public health. Because of this, it´s notable that the increase of urbanization in the last 20 years put in quest all the control strategies existent, the measures implanted until today were incapable to eliminate the transmission as to impede the occurrence of new epidemics. The prevention of dogs through imunoprophylaxis appear as an alternative, however to have a safe use it´s needed other studies, that can bring real results. The research below shows the increase of visceral leishmaniosis during the years, associated mainly to human cases, besides of dismember the tries used or that will be used with objective of dicrease the spread of leishmaniosis

Interactions of maize bushy stunt phytoplasma with the leafhopper vector, Dalbulus maidis (Delong and Wolcott) (Hemiptera: Cicadellidae) and associated microbiota

Phytoplasmas are bacteria with a persistent propagative transmission by insect vectors that generates direct and indirect interactions among them. In order to understand these interactions for maize bushy stunt phytoplasma (MBSP) and the leafhopper vector Dalbulus maidis (Hemiptera: Cicadellidae), two research lines were addressed. The first one aimed to determine the indirect effects of maize infection by MBSP on some biological and behavioral parameters of the vector, whereas a second line investigated direct interactions of the phytoplasma with D. maidis during its movement through the vector body following acquisition from plants, and associated microbiota. Indirect effects were investigated in choice experiments in which alighting and oviposition preferences by D. maidis were compared on healthy vs. MBSP-infected plants with variable incubation time (diseased plants with early and advanced symptoms, or still asymptomatic). Likewise, indirect effect of MBSP on the D. maidis biology was determined in two life table experiments in which the vector was reared on healthy vs. MBSP-infected plants expressing advanced disease symptoms or still asymptomatic. Choice experiments showed that alighting and oviposition preferences of D. maidis on MBSP-infected plants compared to healthy plants depend on the pathogen incubation period in the plant. The leafhopper preferred MBSP-infected plants over healthy ones during the asymptomatic phase of the disease, but rejected infected plants with advanced symptoms. The vector was able to acquire MBSP from asymptomatic infected plants shortly (3 days) after inoculation, but transmission efficiency increased when acquisition occurred at later stages of the pathogen incubation period (≥14 days) in the source plants and the test plants showed disease symptoms faster. These results suggest that MBSP modulates D. maidis preference for asymptomatic infected plants in the early stages of the crop, allowing rapid spread of this pathogen. Maize infection by the phytoplasma had a neutral effect on most life table parameters of D. maidis; a lower net reproductivity rate (Ro) was observed in the cohort reared on MBSP-infected plants with advanced symptoms, which was compensated to some extent by a higher sexual ratio. MBSP acquisition by all vector nymphal stadia was confirmed by PCR, and the pathogen as detected in both male and female reproductive organs. Concerning direct MBSP-vector interactions, transmission electron microscopy analyses showed phytoplasma-like cells in the midgut lumen, microvilli and epithelial cells, suggesting that MBSP enters the epithelium midgut through the microvilli wall. Within the epithelial cells, mitochondria and bacteria-like cells (possibly endosymbionts) were observed together with masses of phythoplasma-like cells. In the hemocoel, phytoplasma-like cells grouped into a matrix were also observed in association with bacteria-like cells similar to those observed in the midgut epithelium. Similar associations were found in the salivary gland. Interestingly, in-situ hybridization (FISH) technique revealed a variation in diversity and abundance of the microbiota in intestine and salivary glands of D. maidis adults over time after MBSP acquisition from plants. Sulcia sp., Cardinium sp. and eubacteria increased their abundance over time, whereas Rickettsia sp. decreased. The frequent association of the vector microbiota with the phytoplasma in some tissues of D. maidis suggests that endosymbiotic bacteria may play some role in MBSP-vector interactions.

Studies on Brazilian anophelines from the northeast and Amazon regions ...

"References" at end of each study.

Estudo sobre a participação de roedores na cadeia de transmissão de Leishmania infantum (Protozoa: Trypanosomatidae) no Rio Grande do Norte

American visceral leishmaniasis is a zoonosis caused by Leishmania infantum and transmitted by the bite of the sand flies Lutzomia longipalpis.The main domestic reservoir is the dog, while foxes and opposums are the known wild reservoirs. However, identification of natural infections with L. infantum in rodents appears for need of investigating the participation of these rodents how source of infection of the parasite. In the present work the Leishmania infantum infection was investigated in rodents captured in Rio Grande do Norte, aiming at to offer subsidies to the understanding of the epidemic chains of LVA in the State. Thirteen Galea spixii were distributed in four groups, being G1 the group control with four animals and the others, G2, G3 and G4, with three animals each. Those animals were intraperitoneally inoculated with 107 promastigotas of L. infantum and accompanied for, respectively, 30, 90 and 180 days. Weekly the animals were monitored as for the corporal weight and rectal temperature. At the end of each stipulated period the animals were killed. Blood were used for determination of the parameters biochemical and haematological, PCR, ELISA, microscopic examination and cultivation in NNN medium. Liver, spleen and lymph node were used in Giemsa-stained impression and cultivation in NNN medium. Liver and spleen fragments were still used in PCR and histopathological, respectively. At the same time 79 rodents of the species Rattus rattus, Bolomys lasiurus, Oligoryzomys nigripis, Oryzomys subflavus and Trichomys apereoides were captured in the Municipal districts of Brejinho, Campo Grande, Coronel Ezequiel, Passa e Fica and Vázea for identification of natural infection with L. infantum. Evidence of infection was checked by direct examination of Giemsa-stained impression of liver, spleen and blood and culture of these tissues in NNN medium. Antibodies were researched by ELISA. They were not found differences among the weigh corporal final, rectal temperature and biochemical and haematological parameters of the Galea spixii controls and infected. The rectal temperature of the animals varied from 36OC to 40OC. For the first time values of the haematocrit (33,6% to 42,8%), hemoglobin (10,2 to 14,5g/dl), erythrocyts number (4,67x106 to 6,90x106/mm3), total leukocytes (0,9x103 to 9,2x103/mm3), platelets (49x103 to 509x103/mm3) total proteins (1,56 to 6,06 g/dl), albumin (1,34 to 3,05 g/dl) and globulins (0,20 to 3,01 g/dl) of the Galea spixii were determined. The lymphocytes were the most abundant leucocytes. Infection for L. infantum was diagnosed in two animals euthanasied 180 days after the infection. In one of the animals was also identified antibodies anti-Leishmania. The parasite was not found in none of the five other species of rodents captured. Galea spixii are resistant to the infection for L. infantum and they are not good models for the study for visceral leishmaniose, although they can act as infection sources. More studies are necessary to determine the paper of the rodents in the epidemic chain of transmission of the visceral leishmaniose in the State of Rio Grande do Norte

Flutuação populacional e análise faunística de cigarrinhas (Cicadellidae) em pomar de ameixeira em Paranapanema, São Paulo, Brasil.

A produção de ameixa no Brasil é de grande importância econômica. O Estado de São Paulo ocupa a 4ª posição na produção nacional e, assim como os demais estados produtores da fruta, vem enfrentando problemas fitossanitários graves devido a Escaldadura das Folhas da Ameixeira. A doença, causada pela bactéria Xylella fastidiosa, caracteriza-se pela necrose das folhas e secamento dos ramos colonizados pela bactéria, provocando o declínio no vigor e na produção, culminando com a morte da planta. A bactéria X. fastidiosa é transmitida por cigarrinhas (Cicadellidae: Cicadellinae). Este trabalho teve como objetivo identificar as espécies de cigarrinhas (Cicadellidae: Cicadellinae; Gyponinae) presentes em um pomar de ameixeira representativo do município de Paranapanema (São Paulo - Brasil), realizando-se a análise faunística além de comparar métodos de coleta e conhecer a flutuação populacional.

Avaliação do potencial de formigas (Hymenoptera: formicidae) como vetores mecânicos de bactérias do gêneroSstaphylococcus no ambiente hospitalar.

In a hospital environment, these bacteria can be spread by insects such as ants, which are characterized by high adaptability to the urban environment. Staphylococcus is a leading cause of hospital infection. In Europe, Latin America, USA and Canada, the group of coagulase negative staphylococci (CoNS) is the second leading cause of these infections, according to SENTRY (antimicrobial surveillance program- EUA). In this study, we investigated the potential of ants (Hymenoptera: Formicidae) as vehicle mechanics of Staphylococcus bacteria in a public hospital, in Natal-RN. The ants were collected, day and night, from June 2007 to may 2008, in the following sectors: hospitals, laundry, kitchen, blood bank. The ants were identified according to the identification key of Bolton, 1997. For the analysis of staphylococci, the ants were incubated in broth Tryptic Soy Broth (TSB) for 24 hours at 35 º C and then incubated on Mannitol Salt Agar. The typical colonies of staphylococci incubated for 24 hours at 35 ° C in Tryptic Soy Agar for the characterization tests (Gram stain, catalase, susceptibility to bacitracin and free coagulase). The identification of CoNS was performed through biochemical tests: susceptibility to novobiocin, growth under anaerobic conditions, presence of urease, the ornithine decarboxylation and acid production from the sugars mannose, maltose, trehalose, mannitol and xylose. The antimicrobial susceptibility examined by disk-diffusion technique. The technique of Polymerase Chain Reaction was used to confirm the presence of mecA gene and the ability to produce biofilm was verified by testing in vitro using polystyrene inert surface, in samples of resistant staphylococci. Among 440 ants, 85 (19.1%) were carrying coagulase-negative staphylococci (CoNS) of the species Staphylococcus saprophyticus (17), Staphylococcus epidermidis (15), Staphylococcus xylosus (13), Staphylococcus hominis hominis (10), Staphylococcus lugdunensis (10), Staphylococcus warneri (6), Staphylococcus cohnii urealyticum (5), Staphylococcus haemolyticus (3), Staphylococcus simulans (3), Staphylococcus cohnii cohnii (2), and Staphylococcus capitis (1). No Staphylococcus aureus was found. Among the isolates, 30.58% showed resistance to erythromycin. Two samples of CoNS (2.35%), obtained from the ant Tapinoma melanocephalum collected in the post-surgical female ward, S. Hominis hominis and S. lugdunensis harbored the mecA gene and were resistant to multiple antibiotics, and the specie S. hominis hominis even showed to be a biofilm producer. This study proves that ants act as carriers of multidrug-resistant coagulase-negative Staphylococci and biofilm producers and points to the risk of the spreading of pathogenic microorganisms by this insect in the hospital environment

Culicídeos vetores em uma unidade de conservação da caatinga na região do seridó no Rio Grande do Norte aspectos da transmissão de doenças

The scarcity of occurrence records of culicid species, transmitting insects of etiologic agents for arboviruses, protozoan diseases and philarioses, and lack of information about their interactions with the forest environment make the Caatinga one of the most unknown biomes in Brazil. The aim of this study was to identify culicid species in a conservation unit located in a preserved area of the Caatinga and the possible associations with local abiotic factors, as well as characterize the natural and artificial ecotopes present in the Caatinga of Rio Grande do Norte state. The Ecological Station of Serido (ESEC-Seridó) is a Caatinga Conservation Unit located in a rural area of Rio Grande do Norte. The hot and dry climate of the Caatinga, along with a short rainy season, favor the adaptation and resistance of plants, resulting in completely distinct landscapes over the course of the year. The study took place over the course of one year, with monthly collections between 10h-12h, 14h-16h and 18h-20h. Adult mosquitoes were collected in a forest area using a manual suction trap with human bait as attractant at the three times and a Shannon trap for night collections. During the rainy season ovitraps were randomly distributed at different sites. The different capture sites were observed and photographed. Immature and adult insects captured were taken to the laboratory for identification. Of the 5081 insects collected in human-bait and Shannon traps, 75% were Mansonia wilsoni. In addition, Aedeomyia, Aedes, Anopheles, Coquillettidia, Culex, Haemagogus, Mansonia and Psorophora were identified. Of the 92 Haemagogus eggs found, 71 were collected on the banks of a reservoir. Mosquito larvae were found and photographed in an artificial breeding site of a neighboring rural area. Humidity had the greatest influence on culicid density in the Caatinga. Rain was important in the appearance of Haemagogus, Anopheles, Aedes and Psorophora. Temperature had a negative influence on Anopheles triannulatus. The occurence of important vector species of diseases in a little known preserved area of the Caatinga as well as environmental and sociocultural questions are factors that favor the emergence of transmitting insects, making these areas potential subject to the resurgence of diseases

Ecologia de culicídeos vetores de febre amarela silvestre em relação ao microclima na Floresta Nacional de Caxiuanã-Melgaço-PA

Os vetores de febre amarela encontram-se distribuídos, nas Américas, nos gêneros, Haemagogus e Sabethes. Os culicídeos hematófagos têm seu ritmo de atividade hematofágica influenciado por fatores endógenos e fatores exógenos, tais como a resposta aos elementos micro-climáticos. O micro-clima e a estratificação dos hospedeiros são os principais determinantes para a ocupação do nicho ecológico de muitas espécies de culicídeos nas florestas tropicais. O estudo foi desenvolvido na torre de medição micrometeorológica na Floresta Nacional de Caxiuanã, Melgaço, Pará, objetivando analisar a distribuição vertical dos vetores de febre amarela silvestre em relação ao microclima da Floresta densa de terra firme, de julho de 2005 a abril de 2006, no solo em plataformas da torre a 8m, 16m e 30m durante 12 horas diurnas e 12 horas noturnas, utilizando atrativo humano. Em cada uma das alturas encontrava-se disponível termo-higrômetro para a medição da temperatura e umidade relativa do ar e no topo da mesma estava um pluviômetro para a medição da precipitação pluviométrica. Obteve-se 25.498 culicídeos, sendo 1028 pertencentes à Haemagogus e 502 à Sabethes, nas coletas diurnas. As espécies do gênero Sabethes e Haemagogus janthinomys mostraram-se acrodendrófilas, com preferência à realização do repasto a 16m e 30m, enquanto que H. leucocelaenus foi coletado em maior quantidade ao nível do solo. Os meses mais chuvosos apresentaram maior quantidade de vetores de febre amarela silvestre, porém essa influência não foi estatisticamente significativa. Por outro lado houve relação significativa entre temperatura, umidade relativa do ar e número de vetores de febre amarela silvestre.

Análise das possíveis regiões organizadoras nucleolares e da atividade nucleolar em Triatoma melanocephalae T. lenti, importantes vetores da doença de Chagas

Triatoma melanocephalae T. lentisão espécies crípticas de triatomíneos pertencentes ao subcomplexo Brasiliensis. Esses organismos foram agrupados no subcomplexo apenas por caracteres morfológicos e pela disposição geográfica. Sendo assim, estudos citogenéticos são considerados como importantes ferramentas na classificação dos triatomíneos e, com isso, podem auxiliar na criação de um plano de profilaxia da doença. Por meio da técnica citogenética de impregnação por íons prata, foi possível visualizar a atividade nucleolar e as Regiões Organizadoras Nucleolares (RONs) desses insetos. T. melanocephalaapresentou três RONs ativas nos autossomos durante a prófase I. T. lenti apresentou duas RONs ativas nos autossomos durante a prófase I e a metáfase I. Ambas as espécies apresentaram o fenômeno de persistência do material nucleolar encontrado em triatomíneos. Sendo assim, por meio da análise das RONs, foi possível observar que T. lenti, quando comparado com os outros organismos do subcomplexo, apresentou marcações semelhantes à T. tibiamaculata e que T. melanocephalanão apresenta nenhuma relação direta com o subcomplexo. Palavras-chave: Citogenética. Taxonomia. Triatominae. Subcomplexo Brasiliensis. ABSTRACT Analysis of nucleolus organizer regions and nucleolar activity in important vectors of Chagas disease (Triatoma melanocephala and T. lenti) Triatoma melanocephalaand T. lentiare important vectors of Chagas disease. These cryptic species of triatomines are grouped in the subcomplexbrasiliensisdue only to morphological characters and geographical distribution. Cytogenetic studies are important to the classification of insects and can assist in creating a disease prevention plan. The aim of the present study was to determine nucleolar activity and nucleolus organizer regions (NORs) in these insects using the cytogenetic method of silver ion impregnation. T. melanocephalaexhibited three active NORs in autosomes during prophase I. T. lentiexhibited two active NORs in autosomes during prophase I and metaphase I. Both species exhibit the persistent nucleolar material found in triatomines. The analysis of NORs in the present study revealed that T. lenti exhibited labeling similar to that found in T. tibiamaculata, which belongs to the subcomplex, whereas T. melanocephalashows no direct relationship with the subcomplex. Keywords: Cytogenetics. Taxonomy. Triatominae. Brasiliensissubcomplex.

Avaliação do potencial de formigas (Hymenoptera: formicidae) como vetores mecânicos de bactérias do gêneroSstaphylococcus no ambiente hospitalar.

In a hospital environment, these bacteria can be spread by insects such as ants, which are characterized by high adaptability to the urban environment. Staphylococcus is a leading cause of hospital infection. In Europe, Latin America, USA and Canada, the group of coagulase negative staphylococci (CoNS) is the second leading cause of these infections, according to SENTRY (antimicrobial surveillance program- EUA). In this study, we investigated the potential of ants (Hymenoptera: Formicidae) as vehicle mechanics of Staphylococcus bacteria in a public hospital, in Natal-RN. The ants were collected, day and night, from June 2007 to may 2008, in the following sectors: hospitals, laundry, kitchen, blood bank. The ants were identified according to the identification key of Bolton, 1997. For the analysis of staphylococci, the ants were incubated in broth Tryptic Soy Broth (TSB) for 24 hours at 35 º C and then incubated on Mannitol Salt Agar. The typical colonies of staphylococci incubated for 24 hours at 35 ° C in Tryptic Soy Agar for the characterization tests (Gram stain, catalase, susceptibility to bacitracin and free coagulase). The identification of CoNS was performed through biochemical tests: susceptibility to novobiocin, growth under anaerobic conditions, presence of urease, the ornithine decarboxylation and acid production from the sugars mannose, maltose, trehalose, mannitol and xylose. The antimicrobial susceptibility examined by disk-diffusion technique. The technique of Polymerase Chain Reaction was used to confirm the presence of mecA gene and the ability to produce biofilm was verified by testing in vitro using polystyrene inert surface, in samples of resistant staphylococci. Among 440 ants, 85 (19.1%) were carrying coagulase-negative staphylococci (CoNS) of the species Staphylococcus saprophyticus (17), Staphylococcus epidermidis (15), Staphylococcus xylosus (13), Staphylococcus hominis hominis (10), Staphylococcus lugdunensis (10), Staphylococcus warneri (6), Staphylococcus cohnii urealyticum (5), Staphylococcus haemolyticus (3), Staphylococcus simulans (3), Staphylococcus cohnii cohnii (2), and Staphylococcus capitis (1). No Staphylococcus aureus was found. Among the isolates, 30.58% showed resistance to erythromycin. Two samples of CoNS (2.35%), obtained from the ant Tapinoma melanocephalum collected in the post-surgical female ward, S. Hominis hominis and S. lugdunensis harbored the mecA gene and were resistant to multiple antibiotics, and the specie S. hominis hominis even showed to be a biofilm producer. This study proves that ants act as carriers of multidrug-resistant coagulase-negative Staphylococci and biofilm producers and points to the risk of the spreading of pathogenic microorganisms by this insect in the hospital environment