Metaproteomics of complex microbial communities in biogas plants

Production of biogas from agricultural biomass or organic wastes is an important source of renewable energy. Although thousands of biogas plants (BGPs) are operating in Germany, there is still a significant potential to improve yields, e.g. from fibrous substrates. In addition, process stability should be optimized. Besides evaluating technical measures, improving our understanding of microbial communities involved into the biogas process is considered as key issue to achieve both goals. Microscopic and genetic approaches to analyse community composition provide valuable experimental data, but fail to detect presence of enzymes and overall metabolic activity of microbial communities. Therefore, metaproteomics can significantly contribute to elucidate critical steps in the conversion of biomass to methane as it delivers combined functional and phylogenetic data. Although metaproteomics analyses are challenged by sample impurities, sample complexity and redundant protein identification, and are still limited by the availability of genome sequences, recent studies have shown promising results. In the following, the workflow and potential pitfalls for metaproteomics of samples from full-scale BGP are discussed. In addition, the value of metaproteomics to contribute to the further advancement of microbial ecology is evaluated. Finally, synergistic effects expected when metaproteomics is combined with advanced imaging techniques, metagenomics, metatranscriptomics and metabolomics are addressed

A Robust and Universal Metaproteomics Workflow for Research Studies and Routine Diagnostics Within 24 h Using Phenol Extraction, FASP Digest, and the MetaProteomeAnalyzer

The investigation of microbial proteins by mass spectrometry (metaproteomics) is a key technology for simultaneously assessing the taxonomic composition and the functionality of microbial communities in medical, environmental, and biotechnological applications. We present an improved metaproteomics workflow using an updated sample preparation and a new version of the MetaProteomeAnalyzer software for data analysis. High resolution by multidimensional separation (GeLC, MudPIT) was sacrificed to aim at fast analysis of a broad range of different samples in less than 24 h. The improved workflow generated at least two times as many protein identifications than our previous workflow, and a drastic increase of taxonomic and functional annotations. Improvements of all aspects of the workflow, particularly the speed, are first steps toward potential routine clinical diagnostics (i.e., fecal samples) and analysis of technical and environmental samples. The MetaProteomeAnalyzer is provided to the scientific community as a central remote server solution at www.mpa.ovgu.de.Peer Reviewe

Systematic functional analysis of rab GTPases reveals limits of neuronal robustness to environmental challenges in flies

Rab GTPases are molecular switches that regulate membrane trafficking in all cells. Neurons have particular demands on membrane trafficking and express numerous Rab GTPases of unknown function. Here, we report the generation and characterization of molecularly defined null mutants for all 26 rab genes in Drosophila. In flies, all rab genes are expressed in the nervous system where at least half exhibit particularly high levels compared to other tissues. Surprisingly, loss of any of these 13 nervous system-enriched Rabs yielded viable and fertile flies without obvious morphological defects. However, all 13 mutants differentially affected development when challenged with different temperatures, or neuronal function when challenged with continuous stimulation. We identified a synaptic maintenance defect following continuous stimulation for six mutants, including an autophagy-independent role of rab26. The complete mutant collection generated in this study provides a basis for further comprehensive studies of Rab GTPases during development and function in vivo

Faktor Faktor Yang Berhubungan Dengan Perilaku Deteksi Dini Kanker Leher Rahim Metode Inspeksi Visual Asam Asetat Di Puskesmas Candiroto Kabupaten Temanggung

Perilaku deteksi dini kanker leher rahim dengan metode IVA adalah bentuk tindakan pemeriksaan deteksi dini kanker leher rahim dengan metode IVA. Kesadaran perempuan Indonesia untuk melakukan deteksi kanker leher rahim secara teratur masih rendah. Hal ini terlihat dari rendahnya cakupan deteksi dini kanker leher rahim dengan metode Inspeksi Visual Asam Asetat. Untuk tingkat Jawa Tengan hanya 1,08% dari jumlah wanita usia 30-50 tahun, Puskesmas Candiroto sekitar 5%. Tujuan penelitian ini adalah menganalisis faktor yang berhubungan dengan perilaku deteksi dini kanker leher rahim metode Inspeksi Visual Asam Asetat di Puskesmas Candiroto Kabupaten Temanggung. Penelitian ini merupakan penelitian dengan pendekatan cross sectional dengan analisis data uji chi-square dengan taraf signifikansi 95%. Subjek penelitian ini adalah wanita usia subur di wilayakh kerja Puskesmas Candiroto sejumlah 100 orang dengan menggunakan simple random sampling. Hasil uji chi-square ada hubungan antara keterpaparan informasi(p=0,1), dukungan suami (p=0,026), dan dukungan teman (p<0,0001) dengan perilaku deteksi dini kanker leher rahim metode IVA, serta tidak ada hubungan antara umur (p=0,07), pendidikan (p=0,17), pekerjaan (p=0,51), pengetahuan ((p=0,36), sikap (p=0,097), jarak (p=0,1), dukungan tenaga kesehatan (p=0,1) dengan perilaku deteksi dini kanker leher rahim metode IVA. dapat disimpulkan faktor yang berhubungan dengan perilaku deteksi dini kanker leher rahim metode Inspeksi Visual Asam Asetat di Puskesmas Candiroto Kabupaten Temanggung yaitu keterpaparan informasi, dukungan suami, dan dukungan teman. Kata Kunci: kanker leher rahim, deteksi dini, IV

Metaproteome analysis reveals that syntrophy, competition, and phage-host interaction shape microbial communities in biogas plants

Background: In biogas plants, complex microbial communities produce methane and carbon dioxide by anaerobic digestion of biomass. For the characterization of the microbial functional networks, samples of 11 reactors were analyzed using a high-resolution metaproteomics pipeline. Results: Examined methanogenesis archaeal communities were either mixotrophic or strictly hydrogenotrophic in syntrophy with bacterial acetate oxidizers. Mapping of identified metaproteins with process steps described by the Anaerobic Digestion Model 1 confirmed its main assumptions and also proposed some extensions such as syntrophic acetate oxidation or fermentation of alcohols. Results indicate that the microbial communities were shaped by syntrophy as well as competition and phage-host interactions causing cell lysis. For the families Bacillaceae, Enterobacteriaceae, and Clostridiaceae, the number of phages exceeded up to 20-fold the number of host cells. Conclusion: Phage-induced cell lysis might slow down the conversion of substrates to biogas, though, it could support the growth of auxotrophic microbes by cycling of nutrients. © 2019 The Author(s)