RFLP analysis of a PCR-amplified fragment of the 16S rRNA gene as a tool to identify Enterococcus strains

Restriction fragment length polymorphism (RFLP) analysis of a PCR-amplified fragment of the 16S rRNA gene was performed on reference strains belonging to 21 different enterococcal species and on 75 Enterococcus isolates recovered from poultry meat, pasteurised milk and fresh cheese. PCR amplification generated a 275 bp fragment, which was digested with three restriction endonucleases (DdeI, HaeIII, HinfI). The strains were divided into five groups (groups A-E) on the basis of their restriction patterns. Five biochemical tests (arabinose, arginine, manitol, methyl-β-D-glucopyranoside and raffinose) were then performed in addition to RFLP analysis to narrow the identification of enterococcal strains to the species level. PCR-RFLP, in conjunction with the selected biochemical tests, allowed the precise identification of the 21 species of Enterococcus included in the present study. This proposed method is relatively simple and rapid and can be useful as an adjunct tool for accurate identification of Enterococcus.

Cellular characterisation of Candida tropicalis presenting fluconazole-related trailing growth

We assessed fluconazole susceptibility in 52 Candida tropicalis clinical strains using seven antifungal susceptibility methods, including broth microdilution (BMD) [standard M27 A3 (with neutral and acid pH), ATB Fungus 3, Vitek 2 system and flow cytometric analysis] and agar-based methods (disk diffusion and E-test). Trailing growth, detection of cell-associated secreted aspartic proteases (Saps) and morphological and ultrastructural traits of these clinical strains were also examined. The ranges of fluconazole 24 h-minimum inhibitory concentration (MIC) values were similar among all methods. The essential agreement among the methods used for MIC determinations was excellent and all methods categorised all strains as susceptible, except for one strain that showed a minor error. The presence of the trailing effect was assessed by six methods. Trailing positivity was observed for 86.5-100% of the strains. The exception was the BMD-Ac method where trailing growth was not observed. Morphological and ultrastructural alterations were detected in C. tropicalis trailing cells, including mitochondrial swelling and cell walls with irregular shapes. We tested the production of Saps in 13 C. tropicalis strains expressing trailing growth through flow cytometry. Our results showed that all of the C. tropicalis strains up-regulated surface Sap expression after 24 h or 48 h of exposure to fluconazole, which was not observed in untreated yeast strains. We concluded that C. tropicalis strains expressing trailing growth presented some particular features on both biological and ultrastructural levels.

A institucionalização da estratégia como prática nos estudos organizacionais

Neste artigo, analisa-se o nível de institucionalização sob a ótica da Estratégia como Prática (Strategy-As-Practice - SAP), adotando a teoria institucional como perspectiva de análise. Por meio de pesquisa bibliográfica, bibliométrica e sociométrica, analisaram-se 24 estudos publicados no Brasil e 76 no exterior. Os elementos analisados foram: número de artigos publicados em cada ano; obras e autores mais citados; autores que mais publicaram; redes de cooperação entre autores e entre instituições, com o auxílio do software UCINET® 6; abrangência geográfica das parcerias; e enfoques de SAP empregados por meio de análise de conteúdo. Como principais resultados, destacam-se a defasagem entre as primeiras publicações na literatura internacional e na brasileira e os autores mais citados, que são Whittington e Jarzabkowski. A constatação de que 14 diferentes países publicaram artigos sobre SAP aponta sua dispersão geográfica e também seu alinhamento com outras abordagens. No Brasil, apesar do número crescente de artigos publicados e da criação de temas em eventos, ainda há grande espaço para crescimento no número de artigos, nas redes de cooperação e nos enfoques pesquisados.

On the immature stages and natural history of Gonioterma exquisita Duckworth (Lepidoptera, Elachistidae, Stenomatinae)

In the present work the natural history and immature stages of Gonioterma exquisita Duckworth, 1964 of the Brazilian cerrado region are described. The caterpillars are external folivorous feeders and present a diet restricted to pubescent-leaved host plants of the genus Byrsonima Rich. (Malpighiaceae). They are solitary caterpillars and they build a hard individual shelter that looks like a planorbid shell, made from silk and frass, covered with trichomes and silk, where they develop until the last larval instar on the host plant, and remain inside it until the adult emergence on herbaceous stratum. The larval development presented eight instars. Egg, larval head capsules, instar duration, pupa, shelter, and behavior are also described. Diapause is mentioned for the first time in this species.

Ichneumonidae (Hymenoptera) parasitoids of Lepidoptera caterpillars feeding on Croton floribundus Spreng (Euphorbiaceae)

Parasitoids of the family Ichneumonidae (Hymenoptera) were obtained during an inventory of Lepidoptera larvae caught feeding in the wild on Croton floribundus (Euphorbiaceae). The Lepidoptera larvae were collected from host plants along trails inside three preserved forest areas in the Brazilian state of São Paulo. Fifteen different species of Ichneumonidae belonging to five subfamilies (Banchinae, Campopleginae, Cremastinae, Mesochorinae and Metopiinae) were obtained. Seven species of Ichneumonidae were reared from leaf rollers: Meniscomorpha sp. (Banchinae) and Leurus caeruliventris (Cresson) (Metopiinae) from Dichomeris sp. (Gelechiidae); Mesochorus sp.1 (Mesochorinae) [as a parasitoid of Hypomicrogaster sp. (Braconidae, Microgastrinae)], Campoplex sp. (Campopleginae) and Leurus sp. from Olethreutinae sp. (Tortricidae); Sphelodon annulicornis Morley (Banchinae) and Eutanygaster brevipennis Cameron (Cremastinae) were also reared from two unidentified species of Gelechiidae. The other eight species were reared from the larvae of exposed feeders: Diradops sp. (Banchinae) from Miselia albipuncta Hampson (Noctuidae), Casinaria sp. (Campopleginae) from Hymenomima conia Prout (Geometridae), Charops sp. (Campopleginae) from Bagisara paulensis Schaus (Noctuidae) and Oxydia vesulia (Cramer) (Geometridae), two species of Hyposoter Förster (Campopleginae) from Semaeopus sp. (Geometridae) and H. conia, two species of Microcharops Roman (Campopleginae) from B. paulensis and an unidentified species of Limacodidae and Mesochorus sp. 2 [reared from what was probably Aleiodes sp. (Braconidae, Rogadinae)] from an unidentified species of Noctuidae.

Biomass and abundance of termites in three remnant areas of Atlantic Forest in northeastern Brazil

Termite biomass and abundance were estimated in three remnant areas of Atlantic Forest in northeastern Brazil: Reserva Biológica de Pedra Talhada-RBPT (Alagoas State), Parque Estadual Dois Irmãos-PEDI (Pernambuco State), and Área de Proteção Ambiental Mata do Buraquinho-APMB (Paraíba State). A quantitative sampling of the termite assemblages present on trunks and leaf litter, in nests, and in the soil was undertaken during the months of June and July (rainy season) and November and December (dry season) of 2000. Thirty seven species of termites were encountered and quantified in the three remnant forest areas. The termite biomass varied from 10.09 to 11.30 g (fresh weight)/m² and abundance varied from 4911 to 5663 individuals/m². The subfamily Nasutitermitinae was dominant in terms of the number of species, biomass, and abundance. The soil-feeders were the most abundant at the three sites, while wood-feeders had the largest biomass at least at the PEDI and APMB sites. Among the microhabitats quantified, termites were most frequently encountered in the soil, being most heavily concentrated at depths of 0-20 cm. Two species stood out in terms of their biomass and abundance: Embiratermes parvirostris Constantino, on the soil, and Nasutitermes corniger (Motschulsky), on decomposing trunks and in nests. Estimates of biomass and abundance suggest that termites play an important role in the decomposition of the plant necromass in remnant forest areas of the Atlantic Forest in northeastern Brazil.

Phloem mobility of Boron in two eucalypt clones

Boron deficiency causes large productivity losses in eucalypt stands in extensive areas of the Brazilian Cerrado region, thus understanding B mobility is a key step in selecting genetic materials that will better withstand B limitation. Thus, in this study B mobility was evaluated in two eucalypt clones (68 and 129), under B sufficiency or B deficiency, after foliar application of the 10B isotope tracer to a single mature leaf. Samples of young tissue, mature leaves and roots were collected 0, 1, 5, 12 and 17 days after 10B application. The 10B:11B isotope ratio was determined by HR-ICP-MS. Samples of leaves and xylem sap were collected for the determination of soluble sugars and polyalcohols by ion chromatography. Boron was translocated within eucalypt. Translocation of foliar-applied 10B to the young tissues, mature leaves and roots was higher in clone 129 than in 68. Seventeen days after 10B application to a single mature leaf, between 14 and 18 % of B in the young tissue was originated from foliar B application. In plants with adequate B supply the element was not translocated out of the labeled leaf.

Critical values of nitrogen indices in tomato plants grown in soil and nutrient solution determined by different statistical procedures

The objective of this study was to establish critical values of the N indices, namely soil-plant analysis development (SPAD), petiole sap N-NO3 and organic N in the tomato leaf adjacent to the first cluster (LAC), under soil and nutrient solution conditions, determined by different statistical approaches. Two experiments were conducted in randomized complete block design with four repli-cations. Tomato plants were grown in soil, in 3 L pot, with five N rates (0, 100, 200, 400 and 800 mg kg-1) and in solution at N rates of 0, 4, 8, 12 and 16 mmol L-1. Experiments in nutrient solution and soil were finished at thirty seven and forty two days after transplanting, respectively. At those times, SPAD index and petiole sap N-NO3 were evaluated in the LAC. Then, plants were harvested, separated in leaves and stem, dried at 70ºC, ground and weighted. The organic N was determined in LAC dry matter. Three statistical procedures were used to calculate critical N values. There were accentuated discrepancies for critical values of N indices obtained with plants grown in soil and nutrient solution as well as for different statistical procedures. Critical values of nitrogen indices at all situations are presented.

Characterization of Tomato yellow spot virus, a novel tomato-infecting begomovirus in Brazil

The objective of this work was the biological and molecular characterization of a begomovirus detected in São Joaquim de Bicas, Minas Gerais, Brazil, named TGV-[Bi2], by determining its host range, complete nucleotide sequence and phylogenetic relationships with other begomoviruses. Biological characterization consisted of a host range study using either sap inoculation or particle bombardment as inoculation methods. The yellow spot virus can infect plants in Solanaceae and Amaranthaceae, including economically importat crops as sweet pepper, and weeds as Datura stramonium and Nicotiana silvestris. For the molecular characterization, the full-length genome (DNA-A and DNA-B) was amplified, cloned and completely sequenced. Sequence comparisons and phylogenetic analyses indicated that TGV-[Bi2] constitutes a novel begomovirus species named Tomato yellow spot virus (ToYSV), closely related to Sida mottle virus (SiMoV).

Collembolan community in broad-leaved forests and in conifer stands of Cryptomeria japonica in Central Japan

Collembolan communities in conifer plantations (Japanese cedar, Cryptomeria japonica) and secondary deciduous broad-leaved forests of varying ages were investigated to determine the extent to which forest conversion (broad-leaved to coniferous) affects the species richness and assemblage composition of Collembola in central Japan. Density and total species richness of Collembola not differed between the broad-leaved and cedar forests except immediately after clear-cutting. The amount of forest-floor organic matter was larger in cedar forests and positively correlated with the species richness of detritus feeders. Species richness of fungal feeders and sucking feeders positively correlated with the species richness of forest-floor plants. There was difference in collembolan species composition between the forest types. The age of the forests seemed to have only small importance for the collembolan community, except during the first four years after clear-cutting. The conversion to artificial cedar stands has not reduced the abundance or species richness of collembolan communities, but has affected community composition. Differences in species composition may be related to the ground floras.

Qualidades físicas e químicas do abacaxi comercializado na CEAGESP São Paulo

O presente trabalho buscou conhecer a variação das características físicas e químicas do abacaxi comercializado no Entreposto Terminal de São Paulo (ETSP), da Companhia de Entrepostos e Armazéns Gerais de São Paulo (CEAGESP), no período de setembro de 2005 a março de 2006. As avaliações foram realizadas na CEAGESP e nos laboratórios do Departamento de Agroindústria, Alimentos e Nutrição da Escola Superior de Agricultura "Luiz de Queiroz" - ESALQ/USP. Foram realizadas coletas em intervalos de duas semanas, em três atacadistas, com as amostras recolhidas de forma aleatória, padronizadas em frutos tipo 10 (caixa contendo 10 frutos), compostas de dez frutos por atacadista/origem. As amostras foram coletadas nos lotes oferecidos para comercialização pelos atacadistas. Foram avaliadas as seguintes características físicas e químicas: peso do fruto inteiro, peso da coroa, avaliação visual da coloração da casca, densidade, coloração da polpa, pH, teor de sólidos solúveis (SS), acidez titulável (AT), além da relação SS/AT. Para a análise estatística, foi utilizado o Programa SAS. Pelos resultados, observou-se que, dentre as regiões produtoras de abacaxi, destacaram-se, com relação à regularidade de oferta para os três atacadistas, os pólos de Canápolis ('Smooth Cayenne'), Sapé e Miracema do Tocantins ('Pérola'). As diferentes regiões produtoras de abacaxi 'Pérola' apresentaram uma grande variação em relação ao peso médio dos frutos comercializados, indicando diferenças na tecnologia de produção adotada. Dentre as regiões produtoras de abacaxi, houve variação estatística no teor de sólidos solúveis ao longo do período analisado nos pólos de Canápolis ('Smooth Cayenne') e Itaberaba, Sapé e Miracema do Tocantins ('Pérola'). Dentre os parâmetros químicos analisados, a determinação do teor de sólidos solúveis por amostragem com uso do refratômetro manual, associado à maturação aparente (cor da casca), pode representar um avanço significativo na indicação da qualidade dos frutos comercializados, o que permite concluir ser desnecessário o uso de etefon na fase de pré-colheita.

Advances in research and development of mango industry

World mango production is spread over 100 countries that produce over 34.3 million tons of fruit annually. Eighty percent of this production is based in the top nine producing nations that also consume upward of 90% of their production domestically. One to 2 percent of fruit is traded internationally in to markets in the European Community, USA, Arabian Peninsula and Asia. This paper outlines some of the recent research and development advances in mango breeding and genomics, rootstock development, disease management and harvest technologies that are influencing the production and quality of mango fruit traded domestically and internationally.

Differential responses of nematode communities to soybean genotypes resistant and susceptible to Heterodera glycines race 3

Heterodera glycines and Helicotylenchus dihystera were the two most abundant plant-parasitic nematodes found in two H. glycines race 3-infested fields, Chapadão do Céu, MS and Campo Alegre, MG. These fields had been planted with resistant (R) and susceptible (S) plants to cyst nematodes. In the first field, soybean (Glycine max) FT-Cristalina (S) was susceptible to H. glycines but resistant to H. dihystera, while GOBR93 122243 (R) was just the opposite. In the second field, M-Soy 8400 (R) was more resistant to the spiral nematode than M-Soy8411 (S), but the resistance to the cyst nematode was not different between the two genotypes. The total abundance of nematodes was not different between the susceptible and resistant plants in the two fields, suggesting that H. dihystera and/or bacterial feeders and other trophic groups replaced the reduced abundance of the cyst nematodes in resistant plants. Bacterial feeders acted as a compensatory factor to plant-parasitic nematodes in ecological function. The populations of fungal feeders were higher in GOBR93 122243 (R) than in susceptible FT-Cristalina (S) in Chapadão do Céu, but lower in M-Soy 8400 (R) than in M-Soy 8411 (S) in Campo Alegre. This is being attributed to the different periods of soil samplings that were made at the florescent period in the first field, and at the final growing cycle in the second field. Only four nematodes, H. glycines, H. dihystera, Acrobeles sp. and Panagrolaimus sp. dominated the nematode resistant community GOBR93 122243 (R) in Chapadão do Céu, but dominance was shared by ten genera in Campo Alegre, which explains why the five diversity indexes (S, d, Ds, H' and T) were higher in resistant plants than in susceptible plants in field two.

Transgenic passionfruit expressing RNA derived from Cowpea aphid-borne mosaic virus is resistant to passionfruit woodiness disease

Sixteen transgenic yellow passionfruit (Passiflora spp.) plants (R0) were obtained which express a non-translatable transgenic RNA corresponding to the 3' region of the NIb gene and the 5' region of the CP gene, derived from the genome of a Brazilian isolate of Cowpea aphid-borne mosaic virus (CABMV). The transgenic plants were propagated by stem cuttings and challenged by sap inoculation with isolates CABMV-MG1 and CABMV-PE1. One transgenic plant (TE5-10) was resistant to the isolate CABMV-MG1, but susceptible to CABMV-PE1. The remaining transgenic plants developed systemic symptoms, equal to non-transformed plants, when inoculated with either isolate. The absence of virus in TE5-10 plants was confirmed by indirect ELISA. Transcription analysis of the transgene demonstrated that the TE5-10 plant did not accumulate transgenic mRNA, even before inoculation. After inoculation, viral RNA was only detected in plants inoculated with CABMV-PE1. These results confirm that the transgenic plant TE5-10 is resistant to isolate CABMV-MG1, and suggest that the resistance mechanism is post-transcriptional gene silencing, which is already activated in the transgenic plants before virus inoculation.

Levantamento e avaliação da incidência das doenças da mandioca no estado da Paraíba

Objetivando-se levantar e quantificar a incidência de doenças da mandioca em plantios comerciais no Estado da Paraíba, realizaram-se coletas em 21 municípios de quatro microrregiões do Estado no primeiro semestre de 2011. As doenças detectadas foram mancha parda (Cercosporidium henningsii), mancha branca (Phaeoramularia manihotis), antracnose (Colletotrichum gloeosporioides f. sp. manihotis) e podridão mole (Fusarium oxysporum). A incidência apresentou-se com variação, sendo os maiores índices constatados nos municípios das microrregiões do Curimataú Ocidental, Sapé e Brejo paraibano. A mancha parda ocorreu em todos os municípios avaliados, sendo seguida da mancha branca, cuja porcentagem de plantas atacadas foi inferior. A antracnose foi detectada em cerca de 70% dos municípios avaliados, porém com incidência reduzida. A maior incidência da podridão causada por F. oxysporum foi observada na cidade de Imaculada, microrregião de Serra do Teixeira.