Re-infestation of houses by Triatoma dimidiata after intra-domicile insecticide application in the Yucatán peninsula, Mexico

In most countries, Chagas disease transmission control remains based on domestic insecticide application. We thus evaluated the efficacy of intra-domicile cyfluthrin spraying for the control of Triatoma dimidiata, the only Chagas disease vector in the Yucatán peninsula, Mexico, and monitored potential re-infestation every 15 days for up to 9 months. We found that there was a re-infestation of houses by adult bugs starting 4 months after insecticide application, possibly from sylvatic/peridomicile areas. This points out the need to take into account the potential dispersal of sylvatic/peridomestic adult bugs into the domiciles as well as continuity action for an effective vector control.

A bacterial artificial chromosome library for Biomphalaria glabrata, intermediate snail host of Schistosoma mansoni

To provide a novel resource for analysis of the genome of Biomphalaria glabrata, members of the international Biomphalaria glabrata Genome Initiative (biology.unm.edu/biomphalaria-genome.html), working with the Arizona Genomics Institute (AGI) and supported by the National Human Genome Research Institute (NHGRI), produced a high quality bacterial artificial chromosome (BAC) library. The BB02 strain B. glabrata, a field isolate (Belo Horizonte, Minas Gerais, Brasil) that is susceptible to several strains of Schistosoma mansoni, was selfed for two generations to reduce haplotype diversity in the offspring. High molecular weight DNA was isolated from ovotestes of 40 snails, partially digested with HindIII, and ligated into pAGIBAC1 vector. The resulting B. glabrata BAC library (BG_BBa) consists of 61824 clones (136.3 kb average insert size) and provides 9.05 × coverage of the 931 Mb genome. Probing with single/low copy number genes from B. glabrata and fingerprinting of selected BAC clones indicated that the BAC library sufficiently represents the gene complement. BAC end sequence data (514 reads, 299860 nt) indicated that the genome of B. glabrata contains ~ 63% AT, and disclosed several novel genes, transposable elements, and groups of high frequency sequence elements. This BG_BBa BAC library, available from AGI at cost to the research community, gains in relevance because BB02 strain B. glabrata is targeted whole genome sequencing by NHGRI.

Anti-leishmanial and structure-activity relationship of ring substituted 3-phenyl-1-(1,4-di-N-oxide quinoxalin-2-yl)-2-propen-1-one derivatives

A series of ring substituted 3-phenyl-1-(1,4-di-N-oxide quinoxalin-2-yl)-2-propen-1-one derivatives were synthesized and tested for in vitro leishmanicidal activity against amastigotes of Leishmania amazonensis in axenical cultures and murine infected macrophages. Structure-activity relationships demonstrated the importance of a radical methoxy at position R3', R4' and R5'. (2E)-3-(3,4,5-trimethoxy-phenyl)-1-(3,6,7-trimethyl-1,4-dioxy-quinoxalin-2-yl)-propenone was the most active. Cytotoxicity on macrophages revealed that this product was almost six times more active than toxic.

In vitro and in vivo experimental models for drug screening and development for Chagas disease

Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i) the identification and validation of parasite targets, (ii) compounds to be screened against the targets or the whole parasite and (iii) a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.

The in vivo antimalarial activity of methylene blue combined with pyrimethamine, chloroquine and quinine

The effectiveness of methylene blue (MB) combined with pyrimethamine (PYR), chloroquine (CQ) or quinine (Q) was examined in a classical four-day suppressive test against a causative agent of rodent malaria, Plasmodium berghei. A marked potentiation was observed when MB was administered at a non-curative dose of 15 mg/kg/day in combination with PYR (0.19 mg/kg/day) or Q (25 mg/kg/day). No synergy was found between MB (15 mg/Kg) and CQ (0.75 mg/Kg). Our results suggest that the combination of MB with PYR or Q may improve the efficacy of these currently used antimalarial drugs.

A comparative cross-sectional study on the prevalence and morbidity of schistosomiasis in a community in northeastern Brazil (1979-2010)

A cross-sectional study on the prevalence and morbidity of schistosomiasis was conducted in the main settlement of the municipality of Alhandra, in the southern coastal region of the state of Paraíba, in 2010. The results of this study were compared with the results of a previous study conducted in the same area in 1979. The systematic sampling per family conglomerate included approximately 10% of the resident population in the urban area of Alhandra. Faecal examinations were performed using the Kato-Katz method. The clinical forms of the disease were classified in accordance with FS Barbosa as Type I - intestinal form, Type II - hepatointestinal form and Type III - hepatosplenic form. The prevalence of the infection in 2010 was 10.05%, whereas in 1979 it was 46.6% among untreated patients. The percentages of the three clinical forms in 2010 were as follows: 95.3% Type I, 4.6% Type II and 0% Type III; in 1979, the percentages were 94.4%, 3% and 2.6% for Types I, II and III, respectively. In 1979, 6.07% of the Biomphalaria glabrata specimens (the intermediate host in this area) excreted cercariae, where in 2010 only 1.27% of the specimens caught excreted the parasite.

Molecular epidemiology of Candida albicans and Candida glabrata strains isolated from intensive care unit patients in Poland

Over the last decades, Candida spp have been responsible for an increasing number of infections, especially in patients requiring intensive care. Knowledge of local epidemiology and analysis of the spread of these pathogens is important in understanding and controlling their transmission. The aim of this study was to evaluate the genetic diversity of 31 Candida albicans and 17 Candida glabrata isolates recovered from intensive care unit patients from the tertiary hospital in Krakow between 2011-2012. The strains were typed by random amplified polymorphic DNA (RAPD) polymerase chain reaction using five primers (CD16AS, HP1247, ERIC-2, OPE-3 and OPE-18). The results of the present investigation revealed a high degree of genetic diversity among the isolates. No clonal relationship was found among the C. albicans strains, whereas two C. glabrata isolates were identical. The source of Candida infection appeared to be mostly endogenous; however, the presence of two clonal C. glabrata strains suggested the possibility of cross-transmission of these pathogens. Our study confirmed the high discriminatory power of the RAPD technique in the molecular typing of Candida clinical isolates. This method may be applied to the evaluation of transmission routes of pathogenic fungi on a local level.

Malaria on the Guiana Shield: a review of the situation in French Guiana

In a climate of growing concern that Plasmodium falciparum may be developing a drug resistance to artemisinin derivatives in the Guiana Shield, this review details our current knowledge of malaria and control strategy in one part of the Shield, French Guiana. Local epidemiology, test-treat-track strategy, the state of parasite drug resistance and vector control measures are summarised. Current issues in terms of mobile populations and legislative limitations are also discussed.

Therapeutic switching: from antidermatophytic essential oils to new leishmanicidal products

This study examined whether the antidermatophytic activity of essential oils (EOs) can be used as an indicator for the discovery of active natural products against Leishmania amazonensis. The aerial parts of seven plants were hydrodistilled. Using broth microdilution techniques, the obtained EOs were tested against three strains of dermatophytes (Trichophyton mentagrophytes, Microsporum gypseum and Microsporum canis). To compare the EOs antifungal and antiparasitic effects, the EOs activities against axenic amastigotes of L. amazonensis were concurrently evaluated. For the most promising EOs, their antileishmanial activities against parasites infecting peritoneal macrophages of BALB/c mice were measured. The most interesting antifungal candidates were the EOs from Cymbopogon citratus, Otacanthus azureus and Protium heptaphyllum, whereas O. azureus, Piper hispidum and P. heptaphyllum EOs exhibited the lowest 50% inhibitory concentration (IC50) values against axenic amastigotes, thus revealing a certain correspondence between both activities. The P. hispidum EO was identified as the most promising product in the results from the infected macrophages model (IC50: 4.7 µg/mL, safety index: 8). The most abundant compounds found in this EO were sesquiterpenes, notably curzerene and furanodiene. Eventually, the evaluation of the antidermatophytic activity of EOs appears to be an efficient method for identifying new potential drugs for the treatment of L. amazonensis.

Intrusive versus domiciliated triatomines and the challenge of adapting vector control practices against Chagas disease

Chagas disease prevention remains mostly based on triatomine vector control to reduce or eliminate house infestation with these bugs. The level of adaptation of triatomines to human housing is a key part of vector competence and needs to be precisely evaluated to allow for the design of effective vector control strategies. In this review, we examine how the domiciliation/intrusion level of different triatomine species/populations has been defined and measured and discuss how these concepts may be improved for a better understanding of their ecology and evolution, as well as for the design of more effective control strategies against a large variety of triatomine species. We suggest that a major limitation of current criteria for classifying triatomines into sylvatic, intrusive, domiciliary and domestic species is that these are essentially qualitative and do not rely on quantitative variables measuring population sustainability and fitness in their different habitats. However, such assessments may be derived from further analysis and modelling of field data. Such approaches can shed new light on the domiciliation process of triatomines and may represent a key tool for decision-making and the design of vector control interventions.

Adição de níquel na solução nutritiva para o cultivo de mudas de umbuzeiro

Com o objetivo de avaliar a influência da adição de Ni na solução nutritiva de Hoagland & Arnon sobre o crescimento e a nutrição mineral de mudas de umbuzeiro, realizou-se este trabalho. O delineamento experimental utilizado foi o inteiramente casualizado, com quatro repetições e seis doses de Ni (0; 0,0005; 0,05; 0,1; 0,5 e 1,0 mmol L-1). Observou-se que o Ni, em pequenas concentrações, estimula o crescimento de mudas de umbuzeiro em solução nutritiva; para o cultivo destas em solução nutritiva de Hoagland & Arnon, recomenda-se a adição de 0,03 mmol L-1 de Ni.

Diversidade de bactérias diazotróficas endofíticas associadas a plantas de milho

Bactérias diazotróficas endofíticas são capazes de promover o crescimento do milho por meio da fixação biológica do nitrogênio (FBN) ou pela produção de fitormônios. Neste estudo, objetivou-se caracterizar a diversidade de bactérias diazotróficas endofíticas associadas a plantas de milho em diferentes locais do Rio Grande do Sul, que apresentavam variações de clima e solo. Para isso, foi usado um método baseado na amplificação do gene nifH grupo I, na análise de fragmentos de restrição (PCR-RFLP) e no seqüenciamento dos genes amplificados. Foram calculados os índices de Shannon-Weaver e Equitabilidade para estimar a diversidade dos diazotróficos, bem como a diversidade de nucleotídeos e divergência entre seqüências, para estimar a diversidade genética das comunidades amostradas. Na avaliação da diferenciação entre as comunidades foi utilizado o teste F ST. Foi detectada maior variação entre as comunidades das diferentes regiões do Estado do que dentro das comunidades de cada região avaliada, particularmente entre comunidades provenientes de diferentes tipos de solo, regime pluviométrico e regiões geográficas. O índice de diversidade de Shannon-Weaver indicou diferenças em termos de diversidade de unidades taxonômicas entre as comunidades avaliadas. As comunidades amostradas da região norte do Rio Grande do Sul, que mostrou maior disponibilidade de água e conteúdo de argila, tenderam a apresentar maior diversidade quando comparada às comunidades amostradas na região sul. A análise de Equitabilidade mostrou a dominância de unidades taxonômicas dentro de cada comunidade avaliada, independentemente da região amostrada. Todas as seqüências obtidas foram classificadas como pertencentes ao gene nifH grupo I. Foram obtidas seqüências pertencentes às classes Alfa, Beta e Gama-proteobactéria. Esses resultados demonstraram que existe grande diversidade de bactérias endofíticas fixadoras de N capazes de colonizar o interior de plantas de milho e que as diferentes condições edafoclimáticas estão correlacionadas com a diversidade dos genes nifH.

Concentração do potássio do solo em sistema de integração lavoura-pecuária em plantio direto submetido a intensidades de pastejo

A dinâmica de K em sistemas de integração lavoura-pecuária (ILP) diverge daquelas de outros sistemas de manejo, porque os sistemas ILP são mais complexos e envolvem, além das práticas relacionadas à cultura de interesse econômico, a introdução do animal. Objetivou-se, neste trabalho, avaliar as concentrações do K do solo em um sistema ILP, em plantio direto, com diferentes intensidades de pastejo (aveia-preta + azevém) de bovinos no inverno e a cultura da soja cultivada no verão. O experimento foi iniciado em maio de 2001 em área pertencente à Fazenda do Espinilho, localizada no município de São Miguel das Missões - RS, em Latossolo Vermelho distroférrico que vinha sendo cultivado em plantio direto desde 1991. Os tratamentos constaram de intensidades de pastejo: 10, 20, 30 e 40 cm de altura do pasto e um tratamento sem pastejo, distribuídos num delineamento experimental de blocos ao acaso, com três repetições. Os teores de K disponível eram originalmente altos e assim se mantiveram ao longo do tempo, independentemente do tratamento de pastejo. Em todas as situações houve a formação de um gradiente decrescente de concentração de K a partir da superfície, sendo maior após pastagem que após soja. A ausência do pastejo, apesar de propiciar menor ciclagem de K, resultou em maiores teores do nutriente no solo, em relação às áreas com animais, especialmente aquelas intensamente pastejadas, devido às perdas causadas, provavelmente, pelas excretas.

Soil aggregation in a crop-livestock integration system under no-tillage

Grazing intensities can influence soil aggregation, which can be temporarily and permanently affected. The objective of this study was to evaluate the aggregate stability in water at the end of a soybean cycle and during pasture development in a crop-livestock integration system under no-tillage and grazing intensities. The experiment was initiated in 2001, in a dystrophic Red Latosol, after soybean harvest. Treatments consisted of pasture (black oat + Italian ryegrass) at heights of 10, 20 and 40 cm, grazed by young cattle, and a control (no grazing), followed by soybean cultivation, in a randomized block design. Soil samples were collected at the end of the soybean cycle (May/2007), during animal grazing (September/2007) and at the end of the grazing cycle (November/2007). The grazing period influences aggregate distribution, since in the September sampling (0-5 cm layer), there was a higher proportion of aggregates > 4.76 mm at all grazing intensities. Soil aggregation is higher in no-tillage crop-livestock integration systems in grazed than in ungrazed areas.

Ciclagem e balanço de potássio e produtividade de soja na integração lavoura-pecuária sob semeadura direta

A ciclagem e balanço de nutrientes em sistemas de integração lavoura-pecuária (ILP) são processos que envolvem o componente solo, a planta e o animal, além da fertilização, em que o manejo dos animais, por alterar as condições bióticas e abióticas do solo, altera a velocidade de decomposição dos resíduos. Objetivou-se avaliar a ciclagem e o balanço de K em sistema ILP sob semeadura direta, em função da intensidade de pastejo por bovinos em pastagem, no inverno, e sua relação com a produtividade de soja, cultivada no verão. O experimento foi iniciado em maio de 2001 em Latossolo Vermelho distroférrico, em área que vinha sendo cultivada em semeadura direta desde 1991. Os tratamentos constaram de intensidades de pastejo contínuo, por bovinos jovens, em mistura de azevém + aveia-preta, representados por alturas do pasto: 10, 20, 30 e 40 cm e sem pastejo (testemunha), distribuídos num delineamento experimental de blocos ao acaso, com três repetições, em que se avaliou um ciclo pastagem/soja (safra 2007/08). Na ciclagem, foram considerados os teores de K na pastagem, na soja e nos animais e, no balanço desse nutriente, na lavoura e no solo, foram considerados os seus teores no solo e as entradas, como fertilizante e saídas, nos grãos de soja e no tecido animal. A ciclagem do K aumentou com o aumento da intensidade de pastejo, e o seu balanço no sistema, apesar de negativo, com maior déficit nas áreas com maior intensidade de pastejo, não influenciou a produtividade da soja.