Some aspects of disease management of cherry leaf spot (Blumeriella jaapii) with special reference to pesticide use

In this review, some aspects of disease management of cherry leaf spot (Blumeriella jaapii) are summarised with special referenceto pesticide use. In the first part of the review, we show the non-chemical control approach (e. g. removal of fallen leaves, planting resistantcultivar) against leaf spot. In the second part of the review, the effect of pesticides including fertilizers (urea) and fungicides on cherry leafspot are discussed. Special attention are given to the fungicides of copper, dodine, captafol, captan, benomil, chlorothalonil, steroldemethylation inhibitors (e.g. fenarimol, fenbuconazole, myclobutanil, tebuconazole), and strobilurins about their effectiveness against cherryleaf spot. In the final part of the review, possibilities of cherry leaf spot control are discussed in integrated and organic cherry orchards

Produção de bioferramentas para o estudo da deubiquitinase USP2

Orientador : Prof. Dr. Silvio Marques ZanataDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Biologia Celular e Molecular. Defesa: Curitiba, 12/12/2016Inclui referências : f. 65-70Resumo: A ubiquitinação está envolvida em vários processos no organismo, desde ciclo celular até vias de sinalização de apoptose. Por esse amplo envolvimento, é de extrema importância uma fina regulação que é realizada por enzimas chamadas deubiquitinases (DUBs) que hidrolisam ligações isopeptídicas. Dentre todas as DUBs, USP é a família mais extensa tendo como membro USP2 que possui duas isoformas fruto de processamento alternativo: USP2a (de 69kDa) e USP2b (de 45kDa) que se diferenciam apenas na região N-terminal. Alguns estudos relacionam essas isoformas com a via da p53, miogênese, tumores de próstata, agressividade de tumores de mama e manutenção dos níveis de Na+ e pressão sanguínea. Apesar desses estudos, a distribuição tecidual da USP2 ainda não está completamente estabelecida. A presença de duas isoformas distintas de processamento e a inexistência de anticorpos comerciais que possam distinguir entre as duas isoformas justifica a produção de reagentes confiáveis para o estudo de USP2. Desde modo, o objetivo foi produzir anticorpo monoclonal, isoforma específico, para USP2a e clonar, produzir proteína recombinante em bactérias e superexpressar USP2b em células eucariontes. USP2a foi produzida em E. coli BL21(DE3)STAR fusionada com etiqueta de histidina e purificada por coluna de Ni-NTA. A proteína purificada foi usada para imunizar camundongos balb/c. O soro policlonal desses animais foi testado por western blot para a presença de anticorpo anti-USP2a quanto ao reconhecimento da proteína endógena e superexpressa em linhagens de próstata LNCaP e RWPE-1 tendo dois animais positivos usados para a fusão com mieloma. Os hibridomas após a diluição limitante foram varridos por ELISA chegando-se a um clone 4GD2 que reconheceu USP2a superexpressa em HEK293T e não reconheceu USP2b superexpressa em HEK293T, indicando que obteve-se sucesso na produção de um anticorpo monoclonal isoforma específico para USP2a. Além disso, a isoforma USP2b foi clonada com sucesso nos vetores pET28a(+) e pcDNA3.1(-)6his. Esta isoforma não foi expressa em E. coli mesmo com várias mudanças no sistema, desde cepa bacteriana até temperatura, meio e tempo de expressão, mas foi superexpressa em HEK293T. As ferramentas produzidas serão de extrema importância para estudos de mapeamento tecidual, análise de cinética e busca de novos parceiros moleculares para essas isoformas. Palavras-chave: USP2, USP2a, USP2b, anticorpo monoclonal, proteína recombinante.Abstract: Several physiological processes involve ubiquitylating modification, from cell cycle to apoptosis pathways. Because this involvement has a large range, it is extremely important that this post-translational modification be tightly regulated. Enzymes called deubiquitinases (DUBs) hydrolyze isopeptide bond and have a pivotal role in this regulation. Among all DUBs, the USP is the largest family. USP2, a member of this family, has two isoforms from alternative splicing: USP2a (69kDa) and USP2b (45kDa) and differences rely within their N-terminal regions. These isoforms have related to p53 pathway, myogenesis, prostate tumors, breast tumors aggressiveness and maintenance of Na+ levels and blood pressure. Despite these studies, the tissue distribution of USP2 is not completely established. Absences of commercial antibody that recognize specific isoforms justify the production of reliable reagents to study USP2. Thus, the aims of this study were (1) to express both USP2a and USP2b in heterologous bacterial system; (2) to overexpress USP2b in mammalian cells and (3) to produce monoclonal antibodies specific to USP2a isoform. USP2a was produced in E. coli BL21(DE3)STAR fused with polyhistidine tag and purified by Ni-NTA column. The purified protein was used to immunize Balb/c mice strain. We have tested the polyclonal serum of these animals by western blot to the presence of antibody anti-USP2a that recognize endogenous and overexpressed protein in prostate line LNCaP and RWPE-1. Two animals were positive and employed in fusion procedures. After hybridoma supernatant screenings for anti-USP2a presence by ELISA, we isolated the clone 4GD2, which was stable and still secreting antibodies after several freeze-thaw cycles. Monoclonal antibody 4GD2 recognizes USP2a overexpressed in HEK293T cells and does not cross-react with USP2b, suggesting that MoAb 4GD2 is highly specific. In addition, USP2b was successfully cloned in pET28a(+) and pcDNA3.1(-)6his vectors, both expression vectors for prokaryotic and eukaryotic systems, respectively. We were not able to express successfully USP2b in E. coli under different protocols (e.g. temperature, medium and time parameters changes), however we expressed His-tagged USP2b in HEK293T cells. The produced tools will be extremely important in further studies, such as tissue and temporal USP2 expression mapping, USP2 expression and disappearing kinetics in cell lines and characterization of new molecular partners for this DUB. Key words: USP2, USP2a, USP2b, monoclonal antibody and recombinant protein

Disinfection of Ultrasound Transducers Using Non-Sterile Tissue Paper in Some Low-Cost Private Ultrasound Centres in Nigeria – Implications for Nosocomial Infection Management

Background/Aims: The incidence of disease outbreaks in clinical settings arising from ultrasound examinations is well documented, and is a source of worry. The ultrasound transducers and the coupling gel are potential sources of these infections since they come in direct contact with the patient’s skin. In this study, we examine the efficacy of the widespread practice of the use of plain non-sterile tissue paper in some low cost private ultrasound centres in our locality as a method of disinfecting ultrasound transducers after each use. Its potential impact on nosocomial infection management in clinical practice is also examined. Methods: Swab samples from convex ultrasound transducers before and after transabdominal scanning of three consecutive patients were obtained from 10 different ultrasound centres in urban and rural areas of Enugu state. Ultrasound coupling gel samples were equally obtained, and all samples cultured for bacteria growth which was quantified in colony forming units per ml (cfu/ml) and reported in 1000/ml. Paired sampled t-test was used to check for significance in reduction in bacterial load before and after the transducer was cleaned.Results: Nine different bacterial strains were isolated. Staphylococcus aureus and Klebsiella spp had the highest percentage of occurrence in all centres. Significant bacteria growth was recorded in the morning before the examination, and plain tissue paper significantly reduced the bacteria load in the ultrasound transducer.Conclusion: Even though disinfecting ultrasound transducers with non-sterile plain tissue paper alone is statistically effective and has the potential to minimize nosocomial infection, it is however not clinically effective and hence not advised

Micro area based spatial distribution of apple scab in an organic apple orchard

In this study, the objective was to report a two-year investigation on micro area based spatial distribution of apple scab in an organic apple orchard. Results showed that number of symptomatic plant part ranged between 587 and 623 on leaf and between 46 and 78 on fruit for an individual tree. Number of asymptomatic plant part ranged between 1034 and 1321 on leaf and between 119 and 193 on fruit. Disease incidence ranged between 35 and 40% on leaf and between 27 and 33% on fruit. Disease aggregation index ranged between 0.115 and 0.298 on leaf and between 0.117 and 0.221 on fruit. Three of the four trees showed significant within canopy aggregation of disease for leaf apple scab symptoms in both years. For fruit apple scab, two of the four trees showed significant random patterns in both years

Preliminary study on micro area based spatial distribution of Monilinia fructigena in an organic apple orchard

In this study, we aimed to report a preliminary study on micro area based spatial distribution of Monilinia fructigenain an organic apple orchard. Results showed that number of symptomatic fruit ranged between 22 and 42 in 2013 and between25 and 35 in 2014. Number of asymptomatic fruit ranged between 111 and 187 in 2013 and between 119 and 167 in 2014.Disease incidence of fruit ranged between 19.7 and 23.2% in 2013 and between 19.1 and 26.5% in 2014. Disease aggregationindex ranged between 0.111 and 0.335 in 2013 and between 123 and 401 in 2014. Three of the four trees showed significantwithin canopy aggregation of disease for fruit brown rot symptoms in both years. However, the remaining one tree exhibitedrandom patterns during both years. Disease aggregation indicated a disease spread by fruit-to-fruit contact and/or an aggregatedpattern of insect damage

Some aspects of disease management of cherry leaf spot (Blumeriella jaapii) with special reference to pesticide use

In this review, some aspects of disease management of cherry leaf spot (Blumeriella jaapii) are summarised with special reference to pesticide use. In the first part of the review, we show the non-chemical control approach (e. g. removal of fallen leaves, planting resistant cultivar) against leaf spot. In the second part of the review, the effect of pesticides including fertilizers (urea) and fungicides on cherry leaf spot are discussed. Special attention are given to the fungicides of copper, dodine, captafol, captan, benomil, chlorothalonil, sterol demethylation inhibitors (e.g. fenarimol, fenbuconazole, myclobutanil, tebuconazole), and strobilurins about their effectiveness against cherry leaf spot. In the final part of the review, possibilities of cherry leaf spot control are discussed in integrated and organic cherry orchards

Farm-scale evaluation of two widely used sanitation practices for reducing primary inoculums of apple scab in organic apple production

Effects of two widely used sanitation practices were evaluated at farm-scale level on leaf degradation and primary infection by Venturia inaequalis in an organic apple orchard (Eperjeske) on two apple cultivars (Jonathan and Prima) from 2011 to 2013. The tested sanitation practices were eradication of fallen leaves by collection and disc cultivation. Treatments of eradication of fallen leaves by collection and disc cultivation reduced signifi cantly (P< 0.001) leaf litter density with 70–85 and 40–55%, respectively, compared to untreated plots in both years. Above treatments in the same order reduced spring scab incidence with 40–50, and 10–20%, respectively, compared to untreated plots. Incidence of leaf scab in autumn was not signifi cantly lower (P< 0.05) in the treatments in the years

Preliminary study on micro area based spatial distribution of Monilinia fructigena in an organic apple orchard

In this study, we aimed to report a preliminary study on micro area based spatial distribution of Monilinia fructigena in an organic apple orchard. Results showed that number of symptomatic fruit ranged between 22 and 42 in 2013 and between 25 and 35 in 2014. Number of asymptomatic fruit ranged between 111 and 187 in 2013 and between 119 and 167 in 2014. Disease incidence of fruit ranged between 19.7 and 23.2% in 2013 and between 19.1 and 26.5% in 2014. Disease aggregation index ranged between 0.111 and 0.335 in 2013 and between 123 and 401 in 2014. Three of the four trees showed significant within canopy aggregation of disease for fruit brown rot symptoms in both years. However, the remaining one tree exhibited random patterns during both years. Disease aggregation indicated a disease spread by fruit-to-fruit contact and/or an aggregatedpattern of insect damage

In vitro sensitivity of Monilinia laxa to fungicides approved in integrated and organic production systems

The aim of this study was to test the in vitro sensitivity of two isolates of Monilinia laxa to fungicides approved in integrated andorganic production systems. In vitro efficacy of 7 fungicides (Champion 50 WP, Kocide 2000, Nordox 75 WG, Olajos rézkén, Kumulus S,Rézkén, Rézoxiklorid) and another 6 fungicides (Score 25 EC, Efuzin 500 SC, Systane, Folicur Solo, Zato Plusz, Rovral) approved in organicand integrated production systems, respectively, were tested against brown rot of sour cherry. The three isolates showed differences insensitivity to fungicides. Fungicides (with active ingredients of copper and sulphur) applied in organic production showed relatively highpercent growth capacity of M. laxa. Rézkén and Kocide 2000 showed the highest and Kumilus S the lowest efficacy against brown rot.Fungicides applied in integrated production showed relatively low percent growth capacity of M. laxa. Score 25 EC showed the lowest andRovral and Folicur solso the highest efficacy against M. laxa