Oligo-DNA Custom Macroarray for Monitoring Major Pathogenic and Non-Pathogenic Fungi and Bacteria in the Phyllosphere of Apple Trees

BACKGROUND: To monitor the richness in microbial inhabitants in the phyllosphere of apple trees cultivated under various cultural and environmental conditions, we developed an oligo-DNA macroarray for major pathogenic and non-pathogenic fungi and bacteria inhabiting the phyllosphere of apple trees. METHODS AND FINDINGS: First, we isolated culturable fungi and bacteria from apple orchards by an agar-plate culture method, and detected 32 fungal and 34 bacterial species. Alternaria, Aureobasidium, Cladosporium, Rhodotorula, Cystofilobasidium, and Epicoccum genera were predominant among the fungi, and Bacillus, Pseudomonas, Sphingomonas, Methylobacterium, and Pantoea genera were predominant among the bacteria. Based on the data, we selected 29 major non-pathogenic and 12 phytopathogenic fungi and bacteria as the targets of macroarray. Forty-one species-specific 40-base pair long oligo-DNA sequences were selected from the nucleotide sequences of rDNA-internal transcribed spacer region for fungi and 16S rDNA for bacteria. The oligo-DNAs were fixed on nylon membrane and hybridized with digoxigenin-labeled cRNA probes prepared for each species. All arrays except those for Alternaria, Bacillus, and their related species, were specifically hybridized. The array was sensitive enough to detect 10(3) CFU for Aureobasidium pullulans and Bacillus cereus. Nucleotide sequencing of 100 each of independent fungal rDNA-ITS and bacterial 16S-rDNA sequences from apple tree was in agreement with the macroarray data obtained using the same sample. Finally, we analyzed the richness in the microbial inhabitants in the samples collected from apple trees in four orchards. Major apple pathogens that cause scab, Alternaria blotch, and Marssonina blotch were detected along with several non-phytopathogenic fungal and bacterial inhabitants. CONCLUSIONS: The macroarray technique presented here is a strong tool to monitor the major microbial species and the community structures in the phyllosphere of apple trees and identify key species antagonistic, supportive or co-operative to specific pathogens in the orchard managed under different environmental conditions

Molecular techniques for pathogen identification and fungus detection in the environment

Many species of fungi can cause disease in plants, animals and humans. Accurate and robust detection and quantification of fungi is essential for diagnosis, modeling and surveillance. Also direct detection of fungi enables a deeper understanding of natural microbial communities, particularly as a great many fungi are difficult or impossible to cultivate. In the last decade, effective amplification platforms, probe development and various quantitative PCR technologies have revolutionized research on fungal detection and identification. Examples of the latest technology in fungal detection and differentiation are discussed here

IRS-2 Deficiency Impairs NMDA Receptor-Dependent Long-term Potentiation

The beneficial effects of insulin and insulin-like growth factor I on cognition have been documented in humans and animal models. Conversely, obesity, hyperinsulinemia, and diabetes increase the risk for neurodegenerative disorders including Alzheimer's disease (AD). However, the mechanisms by which insulin regulates synaptic plasticity are not well understood. Here, we report that complete disruption of insulin receptor substrate 2 (Irs2) in mice impairs long-term potentiation (LTP) of synaptic transmission in the hippocampus. Basal synaptic transmission and paired-pulse facilitation were similar between the 2 groups of mice. Induction of LTP by high-frequency conditioning tetanus did not activate postsynaptic N-methyl-D-aspartate (NMDA) receptors in hippocampus slices from Irs2−/− mice, although the expression of NR2A, NR2B, and PSD95 was equivalent to wild-type controls. Activation of Fyn, AKT, and MAPK in response to tetanus stimulation was defective in Irs2−/− mice. Interestingly, IRS2 was phosphorylated during induction of LTP in control mice, revealing a potential new component of the signaling machinery which modulates synaptic plasticity. Given that IRS2 expression is diminished in Type 2 diabetics as well as in AD patients, these data may reveal an explanation for the prevalence of cognitive decline in humans with metabolic disorders by providing a mechanistic link between insulin resistance and impaired synaptic transmission

Erwinia amylovora Novel Plasmid pEI70: Complete Sequence, Biogeography, and Role in Aggressiveness in the Fire Blight Phytopathogen

Comparative genomics of several strains of Erwinia amylovora, a plant pathogenic bacterium causal agent of fire blight disease, revealed that its diversity is primarily attributable to the flexible genome comprised of plasmids. We recently identified and sequenced in full a novel 65.8 kb plasmid, called pEI70. Annotation revealed a lack of known virulence-related genes, but found evidence for a unique integrative conjugative element related to that of other plant and human pathogens. Comparative analyses using BLASTN showed that pEI70 is almost entirely included in plasmid pEB102 from E. billingiae, an epiphytic Erwinia of pome fruits, with sequence identities superior to 98%. A duplex PCR assay was developed to survey the prevalence of plasmid pEI70 and also that of pEA29, which had previously been described in several E. amylovora strains. Plasmid pEI70 was found widely dispersed across Europe with frequencies of 5–92%, but it was absent in E. amylovora analyzed populations from outside of Europe. Restriction analysis and hybridization demonstrated that this plasmid was identical in at least 13 strains. Curing E. amylovora strains of pEI70 reduced their aggressiveness on pear, and introducing pEI70 into low-aggressiveness strains lacking this plasmid increased symptoms development in this host. Discovery of this novel plasmid offers new insights into the biogeography, evolution and virulence determinants in E. amylovora

Haavahoitajan näkemyksiä ravitsemuksesta osana haavahoitoa

Opinnäytetyönä on laadullinen tutkimus, jonka tarkoituksena on tutkia haavahoitajan näkemyksiä ravitsemuksesta osana haavanhoitoa. Aineisto kerättiin avoimilla essee kyselylomakkeilla. Tutkimuskohteenamme on kolme haavahoitajaa. Lähetimme kolme kyselylomaketta, joihin kaikkiin vastattiin. Aineisto analysoitiin käyttäen sisällönanalyysi menetelmää. Teoreettisessa viitekehyksessä käsittelemme haavaa, ja sen paranemisprosessia, aloittaen ihosta. Teoria jatkuu ravitsemuksella, nestetasapainolla ja aliravitsemuksella. Lisäksi avaamme haavahoitajan määritelmän. Haavahoitajilla tarkoitetaan sairaanhoitajaa, terveydenhoitajaa, ensihoitajaa tai kätilöä, jolla on korkeakoulutasoisia haavanhoitoon liittyviä erikoistumisopintoja vähintään 20 opintoviikkoa suoritettuna. Tässä työssä haavahoitajalla tarkoitamme virallisen haavahoitajan nimikkeen omaavaa työntekijää, joka on kouluttautunut ammattiinsa erikoistumisopinnoilla. Olemme tutkimuslupaa anoessamme huomioineet sen mahdollisuuden, ettei tutkimuskohteessamme olisi riittävästi haavahoitajan erikoistutkintoa omaavia henkilöitä, joten olisimme hyväksyneet haavahoitajaksi myös työn ohessa kouluttautuneen haavavastaavan. Tutkimustulosten mukaan ravitsemus on olennainen osa haavanhoitoa ja haavan paranemisprosessia. Tuloksissa käy ilmi, että haavahoitajilla on monipuoliset tiedot ravitsemuksesta, he kehittävät osaamistaan säännöllisesti monin eri tavoin ja he hyödyntävät tietoa muun muassa potilaan ja omaisten ohjauksessa.This study is qualitative and its purpose was to describe wound carers’ views of nutrition as a part of wound treatment. The material was collected with open questionnaires. The target of this study was three wound carers’ from the surgical polyclinic in the local center hospital. We sent out three questionnaires and got back three answers. The material was analysed using contents method. In the theoretical part we discussed skin, the wound and the healing process of the wound. Then we continued with some theory of food, nutrition, balance of hydration and malnutrition. The text also explained the theory of the official title of a wound carer. The title of wound carer can be used by a nurse, puplic health nurse, paramedic or a midwife, who has at least 20 credits of studies in wound care. In this study we used the title of wound carer for a person who had a certified degree in wound care. When we applied the reseach permit from the hospital we took a note of the fact that we would possibly not have enough nurses to investigate who have the official title of wound carer in the surgical policlinic. We decided just in case to include into our reseach also nurses who have specialised in taking care of cuts

Fumigation with acetic acid vapor to control decay of stored apples

Introduction. Apples are potentially subject to blue mold decay caused by Penicillium expansum if stored at 1 °C for three or more months or if wounded during handling. Results from trials with apples contaminated with conidia of P. expansum and fumigated in small chambers with acetic acid (AA) vapor indicated that fruit could be sterilized to reduce decay without effect on fruit quality. The objective of this study was to determine if larger quantities of apples treated with AA vapor would have less decay after storage and/or wounding. It was also important to determine if fumigation would affect apple quality and aroma. Materials and methods. Apple cultivars were harvested at commercial maturity for use in AA fumigation trials. Apples artificially or naturally contaminated with conidia of P. expansum were fumigated with AA vapor in a 1 m3 gas tight chamber at 10 °C for 1 h to 24 h or dipped in 450 μg thiabendazole × L-1 solution. Fruit fumigated in standard wooden or plastic apple boxes, or small wooden bins were either wounded and evaluated for decay after a week at 20 °C or stored at 1 °C for three or more months and evaluated for decay. Then apple quality was assessed. Results. Apples naturally contaminated with Penicillium spp. that had been stored at 1 °C in air storage and treated with AA vapour had 50% less decay than the control fruit. In another experiment, AA fumigation was as effective as thiabendazole in reducing decay. AA fumigation reduced decay of fruit coming out of storage for apples stored for 3 months, and a second AA fumigation reduced infection of wounds on these same apples. AA fumigation before storage did not affect apple quality or vinegar aroma. Discussion. AA fumigation showed great potential for reducing decay in stored apples. It could be used as an organic alternative to synthetic fungicides for control of blue mold decay