Assessing the Cultivability of Bacteria and Fungi from Arable Crop Residues Using Metabarcoding Data As a Reference
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DNA Metabarcoding of Microbial Communities for Healthcare I
Reviews ISSN 1993-6842 (on-line); ISSN 0233-7657 (print) Biopolymers and Cell. 2016. Vol. 32. N 1. P 3–8 doi: http://dx.doi.org/10.7124/bc.000906 UDC 577.25 + 579.61 DNA metabarcoding of microbial communities for healthcare I. Ye. Zaets1, O. V. Podolich1, O. N. Reva2, N. O. Kozyrovska1 1 Institute of Molecular Biology and Genetics, NAS of Ukraine, 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 2 Department of Biochemistry, Bioinformatics and Computational Biology Unit, University of Pretoria Lynnwood road, Hillcrest, Pretoria, South Africa, 0002 [email protected], [email protected], [email protected] High-throughput sequencing allows obtaining DNA barcodes of multiple species of microorganisms from a single environmental sample. Next Generation Sequencing (NGS)-based profiling provides new opportunities to evaluate the human health effect of microbial community members affiliated to probiotics. DNA metabar- coding may serve as a quality control of microbial communities, comprising complex probiotics and other fermented foods. A detailed inventory of complex communities is a pre-requisite of understanding their func- tionality as whole entities that makes it possible to design more effective bio-products by precise replacement of one community member by others. The present paper illustrates how the NGS-based DNA metabarcoding allows profiling of both wild and hybrid multi-microbial communities with the example of a kombucha probi- otic beverage fermented by yeast-bacterial partners. Keywords: DNA metabarcoding, microbial communities, healthcare, -
Metagenomics Approaches for the Detection and Surveillance of Emerging and Recurrent Plant Pathogens
microorganisms Review Metagenomics Approaches for the Detection and Surveillance of Emerging and Recurrent Plant Pathogens Edoardo Piombo 1,2 , Ahmed Abdelfattah 3,4 , Samir Droby 5, Michael Wisniewski 6,7, Davide Spadaro 1,8,* and Leonardo Schena 9 1 Department of Agricultural, Forest and Food Sciences (DISAFA), University of Torino, 10095 Grugliasco, Italy; [email protected] 2 Department of Forest Mycology and Plant Pathology, Uppsala Biocenter, Swedish University of Agricultural Sciences, P.O. Box 7026, 75007 Uppsala, Sweden 3 Institute of Environmental Biotechnology, Graz University of Technology, Petersgasse 12, 8010 Graz, Austria; [email protected] 4 Department of Ecology, Environment and Plant Sciences, University of Stockholm, Svante Arrhenius väg 20A, 11418 Stockholm, Sweden 5 Department of Postharvest Science, Agricultural Research Organization (ARO), The Volcani Center, Rishon LeZion 7505101, Israel; [email protected] 6 U.S. Department of Agriculture—Agricultural Research Service (USDA-ARS), Kearneysville, WV 25430, USA; [email protected] 7 Department of Biological Sciences, Virginia Technical University, Blacksburg, VA 24061, USA 8 AGROINNOVA—Centre of Competence for the Innovation in the Agroenvironmental Sector, University of Torino, 10095 Grugliasco, Italy 9 Department of Agriculture, Università Mediterranea, 89122 Reggio Calabria, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-0116708942 Abstract: Globalization has a dramatic effect on the trade and movement of seeds, fruits and vegeta- bles, with a corresponding increase in economic losses caused by the introduction of transboundary Citation: Piombo, E.; Abdelfattah, A.; plant pathogens. Current diagnostic techniques provide a useful and precise tool to enact surveillance Droby, S.; Wisniewski, M.; Spadaro, protocols regarding specific organisms, but this approach is strictly targeted, while metabarcoding D.; Schena, L. -
Potato Dextrose Agar with Lecithin and Tween 80 (7575)
POTATO DEXTROSE AGAR w/ LECITHIN & TWEEN 80 (7575) Intended Use Potato Dextrose Agar w/ Lecithin & Tween 80 is used for the isolation of fungi from surfaces sanitized with quaternary ammonium compounds. Product Summary and Explanation Potato Dextrose Agar is a general purpose medium for yeasts and molds that can be supplemented with acid or antibiotics to inhibit bacterial growth. The nutritionally rich base (potato infusion) encourages mold sporulation and pigment production in some dermatophytes.1 Potato Dextrose Agar w/ Lecithin & Tween 80 is a modification of Potato Dextrose Agar. The addition of Lecithin and Tween 80 to Potato Dextrose Agar is used to neutralize antiseptics and disinfectants for environmental monitoring and other applications.2 Complete neutralization of disinfectants is important. Disinfectant carryover can cause a false no-growth test result. Principles of the Procedure Potato Infusion provides nitrogen and vitamin sources required for organism growth. Dextrose is included as a carbon source. Lecithin neutralizes quaternary ammonium compounds and ethanol, and Tween 80 neutralizes phenols, hexachlorophene, and formalin. Agar is the solidifying agent. Formula / Liter Potato Infusion (dehydrated) .................................................... 4 g Dextrose ................................................................................. 20 g Tween 80 .................................................................................. 5 g Lecithin.................................................................................. -
Cristescu TREE 2014.Pdf
TREE-1853; No. of Pages 6 Opinion From barcoding single individuals to metabarcoding biological communities: towards an integrative approach to the study of global biodiversity Melania E. Cristescu Department of Biology, McGill University, Montreal, QC H3A 1B1, Canada DNA-based species identification, known as barcoding, introduced by Arnot et al. [6] and was firmly advanced transformed the traditional approach to the study of and standardized by Hebert et al. [7]. The simple idea of biodiversity science. The field is transitioning from bar- using a short DNA fragment as a barcode (see Glossary) for coding individuals to metabarcoding communities. This identifying species across the Metazoa has been both revolution involves new sequencing technologies, bio- strongly embraced and vigorously scrutinized over the past informatics pipelines, computational infrastructure, and decade [8,9]. Nevertheless, the efforts led by Paul Hebert, experimental designs. In this dynamic genomics land- and supported by the Consortium for the Barcode of Life scape, metabarcoding studies remain insular and biodi- (CBoL; http://www.barcodeoflife.org/) resulted in a global versity estimates depend on the particular methods enterprise that combined molecular tools with valuable but used. In this opinion article, I discuss the need for a scarce taxonomic expertise [10,11]. Today, DNA barcodes coordinated advancement of DNA-based species identi- are being used commonly to identify specimens and the fication that integrates taxonomic and barcoding infor- approach has wide applications in biodiversity conserva- mation. Such an approach would facilitate access to tion, environmental management, invasion biology, the almost 3 centuries of taxonomic knowledge and 1 de- study of trophic interactions, and food safety [12–14]. -
Edna in a Bottleneck: Obstacles to Fish Metabarcoding Studies in Megadiverse Freshwater 3 Systems 4 5 Authors: 6 Jake M
bioRxiv preprint doi: https://doi.org/10.1101/2021.01.05.425493; this version posted January 7, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. 1 Title: 2 eDNA in a bottleneck: obstacles to fish metabarcoding studies in megadiverse freshwater 3 systems 4 5 Authors: 6 Jake M. Jackman1, Chiara Benvenuto1, Ilaria Coscia1, Cintia Oliveira Carvalho2, Jonathan S. 7 Ready2, Jean P. Boubli1, William E. Magnusson3, Allan D. McDevitt1* and Naiara Guimarães 8 Sales1,4* 9 10 Addresses: 11 1Environment and Ecosystem Research Centre, School of Science, Engineering and Environment, 12 University of Salford, Salford, M5 4WT, UK 13 2Centro de Estudos Avançados de Biodiversidade, Instituto de Ciências Biológicas, Universidade 14 Federal do Pará, Belém, Brazil 15 3Coordenação de Biodiversidade, Instituto Nacional de Pesquisas da Amazônia, Manaus, 16 Amazonas, Brazil 17 4CESAM - Centre for Environmental and Marine Studies, Departamento de Biologia Animal, 18 Faculdade de Ciências da Universidade de Lisboa, Lisbon, Portugal 19 20 *Corresponding authors: 21 Naiara Guimarães Sales, [email protected] 22 Allan McDevitt, [email protected] 23 24 Running title: Obstacles to eDNA surveys in megadiverse systems 25 26 Keywords: Amazon, barcoding gap, freshwater, MiFish, Neotropics, reference database, 27 taxonomic resolution 28 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.01.05.425493; this version posted January 7, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. -
Metabarcoding in the Abyss: Uncovering Deep-Sea Biodiversity Through Environmental
Metabarcoding in the abyss : uncovering deep-sea biodiversity through environmental DNA Miriam Isabelle Brandt To cite this version: Miriam Isabelle Brandt. Metabarcoding in the abyss : uncovering deep-sea biodiversity through environmental DNA. Agricultural sciences. Université Montpellier, 2020. English. NNT : 2020MONTG033. tel-03197842 HAL Id: tel-03197842 https://tel.archives-ouvertes.fr/tel-03197842 Submitted on 14 Apr 2021 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE POUR OBTENIR LE GRADE DE DOCTEUR DE L’UNIVERSITÉ DE M ONTPELLIER En Sciences de l'Évolution et de la Biodiversité École doctorale GAIA Unité mixte de recherche MARBEC Pourquoi Pas les Abysses ? L’ADN environnemental pour l’étude de la biodiversité des grands fonds marins Metabarcoding in the abyss: uncovering deep - sea biodiversity through environmental DNA Présentée par Miriam Isabelle BRANDT Le 10 juillet 2020 Sous la direction de Sophie ARNAUD-HAOND et Daniela ZEPPILLI Devant le jury composé de Sofie DERYCKE, Senior researcher/Professeur rang A, ILVO, Belgique Rapporteur -
Experimental Quantification of Pollen with DNA Metabarcoding Using
www.nature.com/scientificreports OPEN Experimental quantifcation of pollen with DNA metabarcoding using ITS1 and trnL Sandra Baksay 1*, André Pornon1, Monique Burrus1, Jérôme Mariette2, Christophe Andalo1 & Nathalie Escaravage1 Although the use of metabarcoding to identify taxa in DNA mixtures is widely approved, its reliability in quantifying taxon abundance is still the subject of debate. In this study we investigated the relationships between the amount of pollen grains in mock solutions and the abundance of high- throughput sequence reads and how the relationship was afected by the pollen counting methodology, the number of PCR cycles, the type of markers and plant species whose pollen grains have diferent characteristics. We found a signifcant positive relationship between the number of DNA sequences and the number of pollen grains in the mock solutions. However, better relationships were obtained with light microscopy as a pollen grain counting method compared with fow cytometry, with the chloroplastic trnL marker compared with ribosomal ITS1 and with 30 when compared with 25 or 35 PCR cycles. We provide a list of recommendations to improve pollen quantifcation. Environmental DNA metabarcoding is a molecular method that consists of investigating environmental DNA samples made of complex mixtures of genomes from numerous organisms1. Due to new sequencing technologies and bioinformatics tools, metabarcoding has been increasingly used to identify taxa in environmental samples1 to monitor biodiversity2–4, to investigate ecosystem functioning5 and interaction networks6–8, in both aquatic and terrestrial ecosystems. Nevertheless, its reliability in quantitative approaches, which depend on the match between counts of high-throughput sequence reads and the amount of sampled biological material2, is still the subject of debate9,10. -
Characterization of Bacterial Communities Associated
www.nature.com/scientificreports OPEN Characterization of bacterial communities associated with blood‑fed and starved tropical bed bugs, Cimex hemipterus (F.) (Hemiptera): a high throughput metabarcoding analysis Li Lim & Abdul Hafz Ab Majid* With the development of new metagenomic techniques, the microbial community structure of common bed bugs, Cimex lectularius, is well‑studied, while information regarding the constituents of the bacterial communities associated with tropical bed bugs, Cimex hemipterus, is lacking. In this study, the bacteria communities in the blood‑fed and starved tropical bed bugs were analysed and characterized by amplifying the v3‑v4 hypervariable region of the 16S rRNA gene region, followed by MiSeq Illumina sequencing. Across all samples, Proteobacteria made up more than 99% of the microbial community. An alpha‑proteobacterium Wolbachia and gamma‑proteobacterium, including Dickeya chrysanthemi and Pseudomonas, were the dominant OTUs at the genus level. Although the dominant OTUs of bacterial communities of blood‑fed and starved bed bugs were the same, bacterial genera present in lower numbers were varied. The bacteria load in starved bed bugs was also higher than blood‑fed bed bugs. Cimex hemipterus Fabricus (Hemiptera), also known as tropical bed bugs, is an obligate blood-feeding insect throughout their entire developmental cycle, has made a recent resurgence probably due to increased worldwide travel, climate change, and resistance to insecticides1–3. Distribution of tropical bed bugs is inclined to tropical regions, and infestation usually occurs in human dwellings such as dormitories and hotels 1,2. Bed bugs are a nuisance pest to humans as people that are bitten by this insect may experience allergic reactions, iron defciency, and secondary bacterial infection from bite sores4,5. -
Introduction to Bioinformatics Analysis of Metabarcoding Data
Introduction to Bioinformatics analysis of Metabarcoding data Theoretical part Alvaro Sebastián Yagüe www.sixthresearcher.com @SixthResearcher • Experimental design • Sampling • Sample processing • Sequencing • Sequence processing www.sixthresearcher.com @SixthResearcher • Experimental design • Sampling • Sample processing • Sequencing • Sequence processing www.sixthresearcher.com @SixthResearcher What do we want to sequence? Whole genome sequencing (WGS) ChIP-seq Amplicon sequencing (AS) Whole exome sequencing (WES) RNA-seq Mass spectrometry www.sixthresearcher.com @SixthResearcher How do we want to sequence? Green,E.D. (2001) Strategies for the systematic sequencing of complex www.sixthresearcher.com genomes. Nat. Rev. Genet., 2, 573–583. @SixthResearcher Metagenomics - Shotgun sequencing www.sixthresearcher.com @SixthResearcher Metagenomics - High-throughput sequencing DNA extraction DNA fragmentation and sequencing Read mapping to reference genome De novo assembly Chimeras! www.sixthresearcher.com @SixthResearcher Metabarcoding - Amplicon sequencing 1. PCR amplification and sample tagging Barcode 1 Barcode 2 Barcode 3 2. Sequencing of PCR products 3. De-multiplexing of reads Samples 1 2 3 4 5 6 Barcode 1 Barcode 2 Barcode 3 www.sixthresearcher.com @SixthResearcher Metabarcoding vs Metagenomics Kress, W. J., & Erickson, D. L. (2008). DNA barcodes: genes, genomics, and bioinformatics. Proceedings of the National Academy of Sciences of the United States of America, 105(8), 2761–2. www.sixthresearcher.com @SixthResearcher Metabarcoding vs Metagenomics http://luckylion.de/2015/10/11/dna-metabarcoding-vs-metagenomics/ www.sixthresearcher.com @SixthResearcher • Experimental design • Sampling • Sample processing • Sequencing • Sequence processing www.sixthresearcher.com @SixthResearcher ➢ Where? Variability in abundance within soil and plant. Consider vertical and horizontal distribution of fungi. ➢ When? Temporal dynamics over short and long term. For complete community census, sample across multiple seasons. -
Environmental DNA Metabarcoding for Simultaneous Monitoring and Ecological Assessment of Many Harmful Algae
bioRxiv preprint doi: https://doi.org/10.1101/2020.10.01.322941; this version posted October 2, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Environmental DNA Metabarcoding for Simultaneous Monitoring and Ecological Assessment of Many Harmful Algae Emily Jacobs-Palmer1, Ramón Gallego1,2, Kelly Cribari1, Abigail Keller1, Ryan P. Kelly1 1University of Washington School of Marine and Environmental Affairs 2NRC Research Associateship Program, Northwest Fisheries Science Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration. Abstract Harmful algae can have profound economic, environmental, and social consequences. As the timing, frequency, and severity of harmful algal blooms (HABs) change alongside global climate, efficient tools to monitor and understand the current ecological context of these taxa are increasingly important. Here we employ environmental DNA metabarcoding to identify patterns in a wide variety of harmful algae and associated ecological communities in the Hood Canal of Puget Sound in Washington State, USA. We track trends of presence and abundance in a series of water samples across nearly two years. We find putative harmful algal sequences in a majority of samples, suggesting that these groups are routinely present in local waters. We report patterns in variants of the economically important genus Pseudo-nitzschia (family Bacillariaceae), as well as multiple harmful algal taxa previously unknown or poorly documented in the region, including a cold-water variant from the saxitoxin-producing genus Alexandrium (family Gonyaulacaceae), two variants from the karlotoxin-producing genus Karlodinium (family Kareniaceae), and one variant from the parasitic genus Hematodinium (family Syndiniaceae). -
Food Microbiology
Food Microbiology Food Water Dairy Beverage Online Ordering Available Food, Water, Dairy, & Beverage Microbiology Table of Contents 1 Environmental Monitoring Contact Plates 3 Petri Plates 3 Culture Media for Air Sampling 4 Environmental Sampling Boot Swabs 6 Environmental Testing Swabs 8 Surface Sanitizers 8 Hand Sanitation 9 Sample Preparation - Dilution Vials 10 Compact Dry™ 12 HardyCHROM™ Chromogenic Culture Media 15 Prepared Media 24 Agar Plates for Membrane Filtration 26 CRITERION™ Dehydrated Culture Media 28 Pathogen Detection Environmental With Monitoring Contact Plates Baird Parker Agar Friction Lid For the selective isolation and enumeration of coagulase-positive staphylococci (Staphylococcus aureus) on environmental surfaces. HardyCHROM™ ECC 15x60mm contact plate, A chromogenic medium for the detection, 10/pk ................................................................................ 89407-364 differentiation, and enumeration of Escherichia coli and other coliforms from environmental surfaces (E. coli D/E Neutralizing Agar turns blue, coliforms turn red). For the enumeration of environmental organisms. 15x60mm plate contact plate, The media is able to neutralize most antiseptics 10/pk ................................................................................ 89407-354 and disinfectants that may inhibit the growth of environmental organisms. Malt Extract 15x60mm contact plate, Malt Extract is recommended for the cultivation and 10/pk ................................................................................89407-482 -
Effects of Growth Media on the Diversity of Culturable Fungi from Lichens
molecules Article Effects of Growth Media on the Diversity of Culturable Fungi from Lichens Lucia Muggia 1,*,†, Theodora Kopun 2,† and Martin Grube 2 1 Department of Life Sciences, University of Trieste, via Giorgieri 10, 34127 Trieste, Italy 2 Institute of Plant Science, Karl-Franzens University of Graz, Holteigasse 6, 8010 Graz, Austria; [email protected] (T.K.); [email protected] (M.G.) * Correspondence: [email protected] or [email protected]; Tel.: +39-04-0558-8825 † These authors contributed equally to the work. Academic Editor: Joel Boustie Received: 1 March 2017; Accepted: 11 May 2017; Published: 17 May 2017 Abstract: Microscopic and molecular studies suggest that lichen symbioses contain a plethora of associated fungi. These are potential producers of novel bioactive compounds, but strains isolated on standard media usually represent only a minor subset of these fungi. By using various in vitro growth conditions we are able to modulate and extend the fraction of culturable lichen-associated fungi. We observed that the presence of iron, glucose, magnesium and potassium in growth media is essential for the successful isolation of members from different taxonomic groups. According to sequence data, most isolates besides the lichen mycobionts belong to the classes Dothideomycetes and Eurotiomycetes. With our approach we can further explore the hidden fungal diversity in lichens to assist in the search of novel compounds. Keywords: Dothideomycetes; Eurotiomycetes; Leotiomycetes; nuclear ribosomal subunits DNA; nutrients; Sordariomycetes 1. Introduction Lichens are self-sustaining symbiotic associations of specialized fungi (the mycobionts), and green algae or cyanobacteria (the photobionts), which are located extracellularly within a matrix of fungal hyphae and from which the fungi derive carbon nutrition [1].