Agriculture and Agri-Food Canada View Institution's Website 7 articles published in JoVE Chemistry Natural Product Discovery with LC-MS/MS Diagnostic Fragmentation Filtering: Application for Microcystin Analysis David R. McMullin1, Shawn Hoogstra2, Kimberlynn P. McDonald1, Mark W. Sumarah1,2, Justin B. Renaud2 1Department of Chemistry, Carleton University Ottawa, 2London Research and Development Center, Agriculture and Agri-Food Canada Diagnostic fragmentation filtering, implemented into MZmine, is an elegant, post-acquisition approach to screen LC-MS/MS datasets for entire classes of both known and unknown natural products. This tool searches MS/MS spectra for product ions and/or neutral losses that the analyst has defined as being diagnostic for the entire class of compounds. Biology A Hydroponic Co-cultivation System for Simultaneous and Systematic Analysis of Plant/Microbe Molecular Interactions and Signaling Naeem Nathoo1,2, Mark A. Bernards2, Jacqueline MacDonald3, Ze-Chun Yuan1,3 1London Research and Development Centre, Agriculture & Agri-Food Canada, 2Department of Biology, University of Western Ontario, 3Department of Microbiology and Immunology, University of Western Ontario The described hydroponic cocultivation system supports intact plants with metal mesh screens and cocultivates them with bacteria. Plant tissue, bacteria, and secreted molecules can then be separately harvested for downstream analyses, simultaneously allowing for the molecular responses of both plant hosts and interacting microbes or microbiomes to be investigated. Biology Introducing an Angle Adjustable Cutting Box for Analyzing Slice Shear Force in Meat Tom Whitesell1, Carmen Avilés2, Jennifer L. Aalhus1, Chris R. Calkins3, Ivy L. Larsen1, Manuel Juárez1 1Lacombe Research Centre, Agriculture and Agri-Food Canada, 2Grupo de investigación MERAGEM, Universidad de Córdoba, 3Department of Animal Science, University of Nebraska Slice shear force is a reference method for beef texture analysis. Using an angle adjustable cutting box could increase its accuracy for research purposes. The results from different locations within the longissimus muscle show a high correlation with Warner-Bratzler shear force methodology and high potential adaptability for different muscles. Immunology and Infection Multiplex Detection of Bacteria in Complex Clinical and Environmental Samples using Oligonucleotide-coupled Fluorescent Microspheres Tim J. Dumonceaux1, Jennifer R. Town1, Janet E. Hill2, Bonnie L. Chaban2, Sean M. Hemmingsen3 1Saskatoon Research Centre, Agriculture and Agri-Food Canada, 2Department of Veterinary Microbiology, University of Saskatchewan, 3Plant Biotechnology Institute, National Research Council of Canada We describe a multiplex method for the detection of microorganisms within a sample using oligonucleotide-coupled fluorescent beads. Amplicon from all organisms within a sample is hybridized to a panel of probe-coupled beads. A Luminex or Bio-Plex instrument is used to query each bead for bead type and hybridization signal. Biology Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System Gang Tian1, Qing Lu2, Li Zhang2, Susanne E. Kohalmi1, Yuhai Cui2 1Department of Biology, University of Western Ontario, 2Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada We have developed a technique to test protein-protein interactions in plant. A yellow fluorescent protein (YFP) is split into two non-overlapping fragments. Each fragment is cloned in-frame to a gene of interest via Gateway system, enabling expression of fusion proteins. Reconstitution of YFP signal only occurs when the inquest proteins interact. Immunology and Infection Biocontained Carcass Composting for Control of Infectious Disease Outbreak in Livestock Tim Reuter1, Weiping Xu2, Trevor W. Alexander1, Brandon H. Gilroyed1, G. Douglas Inglis1, Francis J. Larney1, Kim Stanford3, Tim A. McAllister1 1Agriculture and Agri-Food Canada, Lethbridge Research Centre, 2Department of Bioscience and Biotechnology, Dalian University of Technology, 3Agriculture Centre, Alberta Agriculture and Rural Development Using readily available materials, this biocontained composting system enables effective on-site disposal of large animal carcasses arising in the event of infectious disease outbreak. This procedure kills most infectious agents in carcasses and contaminated manure. Once infectious agent is confirmed non-viable, mature compost can be spread as fertilizer. Biology Catheterization of Intestinal Loops in Ruminants Richard R. E. Uwiera1, John P. Kastelic2, G. Douglas Inglis2 1Department of Agricultural, Food and Nutritional Science, University of Alberta, 2Agriculture and Agri-Food Canada, Lethbridge Research Centre, Lethbridge We describe a novel surgical method for catheterizing 'intestinal loops' within the ileum of sheep. Once animals have recovered from surgery and have cleared antibiotics and analgesics, multiple treatments can be deposited directly in loops via the catheters.