Investigation into in vitro anti-leishmanial combinations of calcium channel blockers and current anti-leishmanial drugs

The need for drug combinations to treat visceral leishmaniasis (VL) arose because of resistance to antimonials, the toxicity of current treatments and the length of the course of therapy. Calcium channel blockers (CCBs) have shown anti-leishmanial activity; therefore their use in combination with standard drugs could provide new alternatives for the treatment of VL. In this work, in vitro isobolograms of Leishmania (Leishmania) chagasi using promastigotes or intracellular amastigotes were utilised to identify the interactions between five CCBs and the standard drugs pentamidine, amphotericin B and glucantime. The drug interactions were assessed with a fixed ratio isobologram method and the fractional inhibitory concentrations (FICs), sum of FICs (ΣFICs) and the overall mean ΣFIC were calculated for each combination. Graphical isobologram analysis showed that the combination of nimodipine and glucantime was the most promising in amastigotes with an overall mean ΣFIC value of 0.79. Interactions between CCBs and the anti-leishmanial drugs were classified as indifferent according to the overall mean ΣFIC and the isobologram graphic analysis.

Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil

Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.

Current situation and perspectives regarding human Chagas disease in midwestern of the state of Minas Gerais, Brazil

Recognising the importance of Chagas disease in Brazil, Bambuí set up epidemiological surveillance for Chagas disease in 1974 and was the first municipality to do so. To ascertain the current epidemiology of Chagas disease in this municipality, 1.782 blood samples from the general population were analysed; 7.7% of samples were found to be seropositive for Chagas disease. A strong positive correlation between increasing age and Chagas disease was evident in both genders, with the highest prevalence in individuals aged over 60 years. Clinically, the cardiodigestive form of Chagas disease was the most common in these samples. These data confirm the interruption of Trypanosoma cruzi transmission, in parallel with a still important residual morbidity of Chagas disease in the county, thus supporting political decisions that will prioritise epidemiological surveillance and medical treatment of Chagas disease in the coming years.

Correlations between major risk factors and closely related Mycobacterium tuberculosis isolates grouped by three current enotyping procedures: a population-based study in northeast Mexico

The characteristics of tuberculosis (TB) patients related to a chain of recent TB transmissions were investigated. Mycobacterium tuberculosis (MTB) isolates (120) were genotyped using the restriction fragment length polymorphism-IS6110 (R), spacer oligotyping (S) and mycobacterial interspersed repetitive units-variable number of tandem repeats (M) methods. The MTB isolates were clustered and the clusters were grouped according to the similarities of their genotypes. Spearman’s rank correlation coefficients between the groups of MTB isolates with similar genotypes and those patient characteristics indicating a risk for a pulmonary TB (PTB) chain transmission were ana- lysed. The isolates showing similar genotypes were distributed as follows: SMR (5%), SM (12.5%), SR (1.67%), MR (0%), S (46.67%), M (5%) and R (0%). The remaining 35 cases were orphans. SMR exhibited a significant correlation (p < 0.05) with visits to clinics, municipalities and comorbidities (primarily diabetes mellitus). S correlated with drug consumption and M with comorbidities. SMR is needed to identify a social network in metropolitan areas for PTB transmission and S and M are able to detect risk factors as secondary components of a transmission chain of TB.

Current status of Paragonimus and paragonimiasis in Ecuador

A review of national and international publications on paragonimiasis in Ecuador, epidemiological records from the Ministry of Public Health and unpublished research data was conducted to summarise the current status of the parasite/disease. The purpose of the review is to educate physicians, policy-makers and health providers on the status of the disease and to stimulate scientific investigators to conduct further research. Paragonimiasis was first diagnosed in Ecuador 94 years ago and it is endemic to both tropical and subtropical regions in 19 of 24 provinces in the Pacific Coast and Amazon regions. Paragonimus mexicanus is the only known species in the country, with the mollusc Aroapyrgus colombiensis and the crabs Moreirocarcinus emarginatus, Hypolobocera chilensis and Hypolobocera aequatorialis being the primary and secondary intermediate hosts, respectively. Recent studies found P. mexicanus metacercariae in Trichodactylus faxoni crabs of the northern Amazon. Chronic pulmonary paragonimiasis is commonly misdiagnosed and treated as tuberculosis and although studies have demonstrated the efficacy of praziquantel and triclabendazole for the treatment of human infections, neither drug is available in Ecuador. Official data recorded from 1978-2007 indicate an annual incidence of 85.5 cases throughout the 19 provinces, with an estimated 17.2% of the population at risk of infection. There are no current data on the incidence/prevalence of infection, nor is there a national control programme.

Antifungal activity of extracts from Atacama Desert fungi againstParacoccidioides brasiliensis and identification ofAspergillus felis as a promising source of natural bioactive compounds

Fungi of the genus Paracoccidioides are responsible for paracoccidioidomycosis. The occurrence of drug toxicity and relapse in this disease justify the development of new antifungal agents. Compounds extracted from fungal extract have showing antifungal activity. Extracts of 78 fungi isolated from rocks of the Atacama Desert were tested in a microdilution assay against Paracoccidioides brasiliensis Pb18. Approximately 18% (5) of the extracts showed minimum inhibitory concentration (MIC) values≤ 125.0 µg/mL. Among these, extract from the fungus UFMGCB 8030 demonstrated the best results, with an MIC of 15.6 µg/mL. This isolate was identified as Aspergillus felis (by macro and micromorphologies, and internal transcribed spacer, β-tubulin, and ribosomal polymerase II gene analyses) and was grown in five different culture media and extracted with various solvents to optimise its antifungal activity. Potato dextrose agar culture and dichloromethane extraction resulted in an MIC of 1.9 µg/mL against P. brasiliensis and did not show cytotoxicity at the concentrations tested in normal mammalian cell (Vero). This extract was subjected to bioassay-guided fractionation using analytical C18RP-high-performance liquid chromatography (HPLC) and an antifungal assay using P. brasiliensis. Analysis of the active fractions by HPLC-high resolution mass spectrometry allowed us to identify the antifungal agents present in the A. felis extracts cytochalasins. These results reveal the potential of A. felis as a producer of bioactive compounds with antifungal activity.

Usability of computerized nursing process from the ICNP® in intensive care units

OBJECTIVE To analyze the usability of Computerized Nursing Process (CNP) from the ICNP® 1.0 in Intensive Care Units in accordance with the criteria established by the standards of the International Organization for Standardization and the Brazilian Association of Technical Standards of systems. METHOD This is a before-and-after semi-experimental quantitative study, with a sample of 34 participants (nurses, professors and systems programmers), carried out in three Intensive Care Units. RESULTS The evaluated criteria (use, content and interface) showed that CNP has usability criteria, as it integrates a logical data structure, clinical assessment, diagnostics and nursing interventions. CONCLUSION The CNP is a source of information and knowledge that provide nurses with new ways of learning in intensive care, for it is a place that provides complete, comprehensive, and detailed content, supported by current and relevant data and scientific research information for Nursing practices.

Checklist of aedine mosquito species (Diptera, Culicidae, Aedini) occurring in Middle and South America (south of the United States) reflecting current generic and subgeneric status

A list of species of tribe Aedini from Middle and South American, south of the United States, with their current generic combinations is provided. Its purpose is to enable entomologists, public health personnel and mosquito control workers to more quickly become familiar with recent formal taxonomic changes within the tribe.

Local distribution of blackfly (Diptera, Simuliidae) larvae in two adjacent streams: the role of water current velocity in the diversity of blackfly larvae

This study aimed to evaluate the influence of water velocity speed on the local distribution and taxocenosis structure of blackfly larvae. The larvae were collected from two adjacent streams located in the municipality of Angra dos Reis (RJ): Caputera River and one of its tributaries. Riffle litter patches were sampled randomly using a 30 x 30 cm quadrat. Four blackfly species were found: Simulium incrustatum s. l. Lutz, 1910; Simulium (Inaequalium) sp. ; Simulium pertinax s. l. Kollar, 1832 and Simulium subpallidum s. l. Lutz, 1909. Among these species, Simulium pertinax s. l. was clearly associated with higher water current speeds, while Simulium subpallidum s. l. showed association with lower water velocities, and Simulium (Inaequalium) sp. had a relatively constant distribution along the water current gradient.

Insects associated with the composting process of solid urban waste separated at the source

Sarcosaprophagous macroinvertebrates (earthworms, termites and a number of Diptera larvae) enhance changes in the physical and chemical properties of organic matter during degradation and stabilization processes in composting, causing a decrease in the molecular weights of compounds. This activity makes these organisms excellent recyclers of organic matter. This article evaluates the succession of insects associated with the decomposition of solid urban waste separated at the source. The study was carried out in the city of Medellin, Colombia. A total of 11,732 individuals were determined, belonging to the classes Insecta and Arachnida. Species of three orders of Insecta were identified, Diptera, Coleoptera and Hymenoptera. Diptera corresponding to 98.5% of the total, was the most abundant and diverse group, with 16 families (Calliphoridae, Drosophilidae, Psychodidae, Fanniidae, Muscidae, Milichiidae, Ulidiidae, Scatopsidae, Sepsidae, Sphaeroceridae, Heleomyzidae, Stratiomyidae, Syrphidae, Phoridae, Tephritidae and Curtonotidae) followed by Coleoptera with five families (Carabidae, Staphylinidae, Ptiliidae, Hydrophilidae and Phalacaridae). Three stages were observed during the composting process, allowing species associated with each stage to be identified. Other species were also present throughout the whole process. In terms of number of species, Diptera was the most important group observed, particularly Ornidia obesa, considered a highly invasive species, and Hermetia illuscens, both reported as beneficial for decomposition of organic matter.

Interaction between Musca domestica L. and its predator Muscina stabulans (Fallén) (Diptera, Muscidae): effects of prey density and food source abundance

Interaction between Musca domestica L. and its predator Muscina stabulans (Fallén) (Diptera, Muscidae): Effects of prey density and food source abundance. The objective of this work was to evaluate the influence of prey density and food source abundance on the predatory behavior of Muscina stabulans over M. domestica. Three predator/prey proportions were evaluated (1:1, 1:3 and 1:6), using 100 third instar predator larvae against second instar prey larvae. Each proportion was maintained using three different levels of food substrate (25, 50 and 100 g). The experiments were carried out in triplicate in BOD incubators (25ºC, UR 70% ± 10% and 12 h photoperiod). The mortality of the M. domestica larvae was 100% under all conditions, except in the 1:6 predator/prey proportion, at the 50g and 100g food substrate levels, where it was 99.99% and 99.22%, respectively. There was a significant increase in the development period of M. stabulans in relation to the increase in prey density and decrease in quantity of food substrate. An increase in the proportion of individuals and a reduction in the amount of resource slowed down larval development. Muscina stabulans pupal weight was proportional to the increase in prey density and the amount of food substrate. The proportion or the density influenced the survival of M. stabulans, with no difference in relation to the amount of food source and consequently in the interaction of the factors. There was no difference between the 1:1 and 1:3 predator-prey densities, with both differing from the 1:6 density.

Effectiveness of fused magnesium potassium phosphate for marandu grass

The current high price of KCl and great dependence on importation to satisfy the Brazilian demand indicate the need for studies that evaluate the efficiency of other K sources, particularly those based on domestic raw material. For this purpose, a greenhouse experiment was conducted with samples of a sandy clay loam Typic Haplustox, in a completely randomized 4 x 3 x 2 factorial design: four K rates (0, 60, 120, and 180 mg kg-1), three sources (potassium chloride (KCl), fused magnesium potassium phosphate (FMPP) and a mixture of 70 % FMPP + 30 % KCl) and two particle sizes (100 and 60 mesh), with three replications. Potassium fertilization resulted in significant increases in shoot dry matter production and in K concentrations, both in soil and plants. The K source and particle size had no significant effect on the evaluated characteristics. Potassium critical levels in the soil and the shoots were 1.53 mmol c dm-3 and 19.1 g kg-1, respectively.

Sewage sludge as nitrogen and phosphorus source for cane-plant and first ratoon crops

The use of sewage sludge in Brazilian agriculture was regulated by the resolution no. 375 Conama, in 2006. However, there is a lack of research to adequate the mineral N and P fertilizer doses to be applied in agricultural fields treated with this residue. In a field experiment, the effects of application rates of sewage sludge and mineral N and P fertilizers on the productivity and technical characteristics of the cane-plant and first ratoon (residual effect) crops were evaluated. Four doses of sewage sludge (0, 3.6, 7.2 and 10.8 t ha-1, dry base), of N (0, 30, 60 and 90 kg ha-1) and of P2O5 (0, 60, 120 and 180 kg ha-1) were combined in a factorial and laid out on randomized block design, a with two replications. To evaluate the residual effect of the sludge, 120 kg ha-1 N and 140 kg ha-1 of K2O were applied in all plots. Sludge application at cane planting, with or without N and/or P fertilizer increased the stalk yield from 84 up to 118 t ha-1, with no alteration in the sugarcane quality, compared with the application of NPK fertilizer alone, resulting in a stalk yield of 91 t ha-1. The study of the response surface for stalk yield on lowfertility soil was the basis for a recommendation of mineral N and P fertilizer doses for sugarcane implantation as related to sewage sludge application rates. It was also concluded that a sludge application of 10.8 t ha-1, which is the sludge dose established based on the N criterion according to the resolution Conama nº 375, could a) reduce the use of mineral N by 100 % and of P2O5 by 30 %, with increments of 22 % in stalk yield, as a direct effect of sludge application to cane plant crop, and b) increase the stalk yield in the second harvest (first ratoon) by up to 12 % and sugar yield by up to 11 %, by the residual effect of sludge application to sugar cane.

Availability of a soluble phosphorus source applied to soil samples with different acidicity levels

Considerations on the interactions of P in the soil-plant system have a long history, but are still topical and not yet satisfactorily understood. One concern is the effect of liming before or after application of soluble sources on the crop yield and efficiency of available P under these conditions. The aim of this study was to evaluate the effect of soil acidity on availability of P from a soluble source, based on plant growth and chemical extractants. Nine soil samples were incubated with a dose of 200 mg kg-1 P in soil with different levels of previously adjusted acidity (pH H2O 4.5; 5.0; 5.5; 6.0 and 6.5) and compared to soils without P application. After 40 days of soil incubation with a P source, each treatment was limed again so that all pH values were adjusted to 6.5 and then sorghum was planted. After the first and second liming the P levels were determined by the extractants Mehlich-1, Bray-1 and Resin, and the fractionated inorganic P forms. In general, the different acidity levels did not influence the P availability measured by plant growth and P uptake at the studied P dose. For some soils however these values increased or decreased according to the initial soil pH (from 4.5 to 6.5). Plant growth, P uptake and P extractable by Mehlich-1 and Bray-1 were significantly correlated, unlike resin-extractable P, at pH values raised to 6.5. These latter correlations were however significant before the second liming. The P contents extracted by Mehlich-1 and Bray-1 were significantly correlated with each other in the entire test range of soil acidity, even after adjusting pH to 6.5, besides depending on the soil buffering capacity for P. Resin was also sensitive to the properties that express the soil buffering capacity for P, but less clearly than Mehlich-1 and Bray-1. The application of triple superphosphate tended to increase the levels of P-Al, P-Fe and P-Ca and the highest P levels extracted by Bray-1 were due to a higher occurrence of P-Al and P-Fe in the soils.

Ammonium chloride as nitrogen source in sugarcane harvested without burning

The great difficulty of incorporation of N fertilizers into the "green sugarcane" system causes concern and since urea is the most commonly used source, there is the risk of loosing NH3 through volatilization. For this reason, a field experiment was undertaken (in a Hapludox Typic) with the objective of evaluating the agronomic efficiency of ammonium chloride on stubble of the second ratoon (SP89 1115), as well as its residual effect on the subsequent cycle (third ratoon). The experimental design was randomized blocks with four replications. Treatments consisted of three N rates (60, 120 and 180 kg ha-1 N) in the form of NH4Cl, in addition to a control treatment without the addition of N fertilizer. The ratoon cane of the second cutting was harvested in November 2006 and the treatments were applied in December 2006. The second ratoon was harvested mechanically in November 2007 and in December 2007, 450 kg ha-1 of the NPK mixture 20-05-19 was applied, providing 90, 22 and 86 kg ha-1 N, P2O5 and K2O, respectively, for the purpose of evaluating the effect of residual-N from the treatments implanted in December 2006. An increase in the rates of N-NH4Cl had a positive effect on the leaf concentrations of P, Mg and S. Stalk yield (MSS - Mg ha-1 of sugarcane stalks) and sugar (MSH - Mg ha-1 of sucrose) in the November 2006 harvest responded linearly to the increase of N doses in the form of NH4Cl. In relation to the effect of residual-N in the 2007/2008 harvest, it was observed, in general, that the concentrations of macronutrients in the sugarcane leaf +1 were within the range considered adequate in the state of São Paulo, Brazil. The residual-N of the NH4Cl doses resulted in a significant reduction in stalk (MSS) and sugar (MSH) production. It may be concluded that the NH4Cl source at a dose of 120 kg ha-1 N in ratoon fertilization of the second cutting was agronomically efficient, presenting, however, less efficiency of residual-N in the subsequent cycle.