Bioactive endophytic fungi isolated from Caesalpinia echinata Lam. (Brazilwood) and identification of beauvericin as a trypanocidal metabolite from Fusarium sp.

Aiming to identify new sources of bioactive secondary metabolites, we isolated 82 endophytic fungi from stems and barks of the native Brazilian tree Caesalpinia echinata Lam. (Fabaceae). We tested their ethyl acetate extracts in several in vitro assays. The organic extracts from three isolates showed antibacterial activity against Staphylococcus aureus and Escherichia coli [minimal inhibitory concentration (MIC) 32-64 μg/mL]. One isolate inhibited the growth of Salmonella typhimurium (MIC 64 μg/mL) and two isolates inhibited the growth of Klebsiella oxytoca (MIC 64 μg/mL), Candida albicans and Candida tropicalis (MIC 64-128 μg/mL). Fourteen extracts at a concentration of 20 μg/mL showed antitumour activities against human breast cancer and human renal cancer cells, while two isolates showed anti-tumour activities against human melanoma cancer cells. Six extracts were able to reduce the proliferation of human peripheral blood mononuclear cells, indicating some degree of selective toxicity. Four isolates were able to inhibit Leishmania (Leishmania) amazonensis and one isolate inhibited Trypanosoma cruzi by at least 40% at 20 μg/mL. The trypanocidal extract obtained from Fusarium sp. [KF611679] culture was subjected to bioguided fractionation, which revealed beauvericin as the compound responsible for the observed toxicity of Fusarium sp. to T. cruzi. This depsipeptide showed a half maximal inhibitory concentration of 1.9 μg/mL (2.43 μM) in a T. cruzi cellular culture assay.

Molecular identification of Saint Louis encephalitis virus genotype IV in Colombia

Saint Louis encephalitis virus (SLEV) is a member of the Japanese-encephalitis virus serocomplex of the genus Flavivirus. SLEV is broadly distributed in the Americas and the Caribbean Islands, where it is usually transmitted by mosquitoes of the genus Culex and primarily to birds and mammalian-hosts. Humans are occasionally infected by the virus and are dead-end hosts. SLEV causes encephalitis in temperate regions, while in tropical regions of the Americas, several human cases and a wide biological diversity of SLEV-strains have been reported. The phylogenetic analysis of the envelope (E) protein genes indicated eight-genotypes of SLEV with geographic overlap. The present paper describes the genotyping of two SLEV viruses detected in mosquito-pools collected in northern Colombia (department of Cordoba). We used reverse transcription-polymerase chain reaction to amplify a fragment of theE-gene to confirm the virus identity and completeE-gene sequencing for phylogenetic analysis and genotyping of the two-SLEV viruses found circulating in Córdoba. This is the first report of SLEV genotype IV in Colombia (Córdoba) in mosquitoes from a region of human inhabitation, implicating the risk of human disease due to SLEV infection. Physicians should consider SLEV as a possible aetiology for undiagnosed febrile and neurologic syndromes among their patients who report exposure to mosquito-bites.

Identification of the nicotinamide mononucleotide adenylyltransferase of Trypanosoma cruzi

The intracellular parasite Trypanosoma cruzi is the aetiological agent of Chagas disease, a public health concern with an increasing incidence rate. This increase is due, among other reasons, to the parasite’s drug resistance mechanisms, which require nicotinamide adenine dinucleotide (NAD+). Furthermore, this molecule is involved in metabolic and intracellular signalling processes necessary for the survival of T. cruzi throughout its life cycle. NAD+ biosynthesis is performed by de novo and salvage pathways, which converge on the step that is catalysed by the enzyme nicotinamide mononucleotide adenylyltransferase (NMNAT) (enzyme commission number: 2.7.7.1). The identification of the NMNAT of T. cruzi is important for the development of future therapeutic strategies to treat Chagas disease. In this study, a hypothetical open reading frame (ORF) for NMNAT was identified in the genome of T. cruzi. The corresponding putative protein was analysed by simulating structural models. The ORF was amplified from genomic DNA by polymerase chain reaction and was further used for the construction of a corresponding recombinant expression vector. The expressed recombinant protein was partially purified and its activity was evaluated using enzymatic assays. These results comprise the first identification of an NMNAT in T. cruzi using bioinformatics and experimental tools and hence represent the first step to understanding NAD+ metabolism in these parasites.

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.

The identification and differentiation of the Candida parapsilosis complex species by polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer region of the rDNA

Currently, it is accepted that there are three species that were formerly grouped under Candida parapsilosis: C. para- psilosis sensu stricto, Candida orthopsilosis, andCandida metapsilosis. In fact, the antifungal susceptibility profiles and distinct virulence attributes demonstrate the differences in these nosocomial pathogens. An accurate, fast, and economical identification of fungal species has been the main goal in mycology. In the present study, we searched sequences that were available in the GenBank database in order to identify the complete sequence for the internal transcribed spacer (ITS)1-5.8S-ITS2 region, which is comprised of the forward and reverse primers ITS1 and ITS4. Subsequently, an in silico polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to differentiate the C. parapsilosis complex species. Ninety-eight clinical isolates from patients with fungaemia were submitted for analysis, where 59 isolates were identified as C. parapsilosis sensu stricto, 37 were identified as C. orthopsilosis, and two were identified as C. metapsilosis. PCR-RFLP quickly and accurately identified C. parapsilosis complex species, making this method an alternative and routine identification system for use in clinical mycology laboratories.

Ankle-Brachial Index: Nurses Strategy To Cardiovascular Disease Risk Factors Identification

Elevated risk of fatal and non-fatal cardiovascular events is associated with high prevalence of peripheral arterial disease, with assessment through the ankle-brachial index (ABI). This study aimed to demonstrate that the ABI and the Edinburgh Claudication Questionnaire are tools to be used by nurses in prevention and/or treatment of CVD (cardiovascular disease). A cross-sectional study was carried out with patients from a cardiovascular clinic. The Edinburgh Claudication Questionnaire was applied and the ABI was measured with the formula (ABI= Blood Pressure Ankle/Blood Pressure Brachial). A total of 115 patients were included, most were females (57.4%), aged 60.6 ± 12.5 years. The most prevalent risk factors were hypertension (64.3%), physical inactivity (48.7%) and family history (58.3%). The study showed that abnormal ABI was frequently found and 42.6% of the patients with abnormal ABI showed intermittent claudication. The method to evaluate the ABI associated to the Edinburg Claudication Questionnaire, can be easily used by nurses in the clinical evaluation of asymptomatic and symptomatic CVD patients.

Identification of care needs of patients with and without the use of a classification instrument

Objective: To analyze the agreement and disagreement between the assessments by applying or not a patient classification instrument, and to investigate the association between the agreement and personal and professional characteristics of the evaluators. Method: This is a descriptive exploratory study. 105 patients were hospitalized in a teaching hospital in the state of Sao Paulo, using the kappa statistic (weighted) and the Bootstrap method. Results: The agreement between the assessments were​​: kw 0.87 (instrument x internal evaluator), kw 0.78 (instrument x external evaluator) and kw 0.76 (between evaluators) and the influence of some personal and professional characteristics. The assessments conducted through the use of an instrument contemplated a greater number of areas of care in relation to when the instrument was not applied. Conclusion: The use of this instrument is recommended in order to more effectively identify care needs of patients.




Avaliação do curso de gerenciamento online na perspectiva dos egressos

RESUMO Objetivo Avaliar curso online na perspectiva do egresso e verificar a relação entre variáveis. Método Estudo quantitativo, descritivo e exploratório, aplicado aos participantes no final de três versões de um curso de atualização onlinena temática de Gerenciamento em Enfermagem. Resultados Os índices de satisfação nas três categorias elencadas, em três anos, apresentaram resultados acima de 75,0%. Os coeficientes obtidos indicaram alta consistência do questionário. Considerando o índice total, a categoria Desempenho do tutor foi a de índice mais alto. Fortes associações entre Autoavaliação e Desempenho do tutor, Autoavaliação e Programa do curso e Desempenho do tutor e Programa do curso foram identificadas. Não houve associação entre as três categorias referidas com as demais variáveis do estudo. Conclusão Os egressos demonstraram satisfação com o curso, que favoreceu a interação e a promoção do conhecimento coletivo no gerenciamento em enfermagem. Foram reconhecidos, também, aspectos que carecem de melhorias, com destaque à capacitação do tutor para mediar discussões e estimular o envolvimento do estudante ao longo do curso.

Descriptions of new species with a key to identification of the genus Neodexiopsis Malloch (Diptera, Muscidae) in Brazil

Nine new species of Neodexiopsis Malloch from Paraná, southern Brazil, are described: Neodexiopsis cinerea sp. nov. and N. paranaensis sp. nov. from Ponta Grossa; N. facilis sp. nov., N. legitima sp. nov., N. similis sp. nov. and N. uber sp. nov. from Guarapuava; N. erecta sp. nov., N. pura sp. nov. and N. rara sp. nov. from Colombo. A key to the identification of the Brazilian species of Neodexiopsis is also presented.

Taxonomy and morphology of Apsil Malloch (Diptera, Muscidae, Coenosiinae, Coenosiini) with new records, description of a new species and a key to identification

Taxonomy and morphology of Apsil Malloch (Diptera, Muscidae, Coenosiinae, Coenosiini) with new records, description of a new species and a key to identification. Apsil Malloch (Diptera, Muscidae, Coenosiinae, Coenosiini) includes 10 species, most of them described from Chile, but some also from Argentina. Five of them and one new species were found at the California Academy of Sciences collection (San Francisco, California) during the course of a project developed in that institution. Almost all material studied was collected in Chile, mainly by M. E. Irwin, during the year of 1966. Brief diagnosis of the known studied species (A. apicata Malloch, A. atripes Malloch, A. dilata Malloch, A. maculiventris Malloch (female described for the first time) and A. spatulata, Malloch), the description of A. mallochi, sp. nov. and a key for the identification of all known species are given. Color illustrations of some morphological characters make easier the recognition of the species. New geographic records were assigned.

Pictorial identification key for species of Sarcophagidae (Diptera) of potential forensic importance in southern Brazil

Pictorial identification key for species of Sarcophagidae (Diptera) of potential forensic importance in southern Brazil. Species of the subfamily Sarcophaginae are important to forensic entomology due to their necrophagous habits. This contribution presents a pictorial key for the identification of 22 Sarcophaginae species in 10 genera that are commonly found in southern Brazil. Photographs of the main structures used in species identification, mainly from the male terminalia, are provided.

Identification of sap-feeder instars in Angelabella tecomae Vargas & Parra (Lepidoptera, Gracillariidae) feeding on Tecoma fulva fulva (Bignoniaceae)

The number of sap-feeding instars of the leaf-miner Angelabella tecomae Vargas & Parra, 2005 was determined by measuring the head capsule exuviae collected from mines (n = 125) on the shrub Tecoma fulva fulva D. Don (Bignoniaceae) in the Azapa Valley, northern Chile. Sap-feeding larvae went through four instars in 120 occasions while five larvae went through five instars. The possible causes for this variation and the usefulness of this tool for future studies are discussed.

White grubs (Coleoptera, Melolonthidae) in the "Planalto Region", Rio Grande do Sul state, Brazil: Key for identification, species richness and distribution

White grubs (Coleoptera, Melolonthidae) in the "Planalto Region", Rio Grande do Sul state, Brazil: Key for identification, species richness and distribution. The objective of this study was to survey the occurrence and geographic distribution of white grub species (Coleoptera, Melolonthidae) in cultivated and non-cultivated fields of the "Planalto Region", Rio Grande do Sul state, Brazil and develop a key at genus-level. Twenty-eight species from 15 genera and three subfamilies were recorded: Dynastinae, Melolonthinae and Rutelinae. The species or genera recorded for the first time in the state are: Cyclocephala metrica, C. tucumana, Isonychus albicinctus, Liogenys bidenticeps, L. fusca, L. obesa and L. sinuaticeps, Paranomala violacea, as well as unidentified species of Amononyx, Dicrania, Leucothyreus, Macrodactylus, Plectris and Rhizogeniates. Among the species recorded, 23 were associated with winter crops. Only Cyclocephala metrica, Dyscinetus rugifrons, two species of Leucothyreus and one species of the tribe Sericini were not present in cultivated crop fields. Cyclocephala flavipennis and Diloboderus abderus occurred in most of the municipalities sampled, often associated with Plectris sp., C. modesta and C. putrida. The highest richness of melolonthids was concentrated in the northeast of the Planalto region.

Prey identification in nests of the potter wasp Hypodynerus andeus (Packard) (Hymenoptera, Vespidae, Eumeninae) using DNA barcodes

Prey identification in nests of the potter wasp Hypodynerus andeus (Packard) (Hymenoptera, Vespidae, Eumeninae) using DNA barcodes. Geometrid larvae are the only prey known for larvae of the Neotropical potter wasp Hypodynerus andeus (Packard, 1869) (Hymenoptera, Vespidae, Eumeninae) in the coastal valleys of the northern Chilean Atacama Desert. A fragment of the mitochondrial gene cytochrome oxidase c subunit 1 was amplified from geometrid larvae collected from cells of H. andeus in the Azapa Valley, Arica Province, and used to provide taxonomic identifications. Two species, Iridopsis hausmanni Vargas, 2007 and Macaria mirthae Vargas, Parra & Hausmann, 2005 were identified, while three others could be identified only at higher taxonomic levels, because the barcode reference library of geometrid moths is still incomplete for northern Chile.

Comparative morphology and identification key for females of nine Sarcophagidae species (Diptera) with forensic importance in Southern Brazil

ABSTRACTThe identification of female flesh flies was always considered a difficult task since morphological descriptions and keys for females are rare. Even in a forensic entomology framework, where females play a major role, female flesh flies are usually not identified. In order to fill this gap in Southern Brazil fauna we provide detailed descriptions and key for the female of nine species included in four genera: Microcerella halli (Engel), Oxysarcodexia paulistanensis (Mattos), Oxysarcodexia riograndensis (Lopes), Peckia (Euboettcheria) australis (Townsend), Peckia(Euboettcheria) florencioi (Prado and Fonseca), Peckia (Pattonella) intermutans (Walker), Peckia(Pattonella) resona (Lopes), Peckia (Sarcodexia) lambens (Wiedemann), and Sarcophaga(Bercaea) africa (Wiedemann). These species are distinguished mainly by genital characters as tergite 6 divided or undivided, presence of tergite 8, spermatheca morphology and vaginal plate shape.