United States Department of Agriculture 12 articles published in JoVE Environment Sampling for Estimating Frankliniella Species Flower Thrips and Orius Species Predators in Field Experiments Joseph Funderburk1, Xavier Martini1, Josh Freeman1, Iris Strzyzewski1, Edward Traczyk1, Thomas Skarlinsky2, Scott Adkins3 1North Florida Research and Education Center, University of Florida, 2Animal and Plant Health Inspection Services, Plant Protection Quarantine, Miami Plant Inspection Station, United States Department of Agriculture, 3Agricultural Research Service, US Horticultural Research Laboratory, United States Department of Agriculture Presented here is a protocol to determine the number of thrips and minute pirate bug predators in crops over multiple dates in field experiments. Also illustrated is how to determine the efficacy of management tactics against thrips and evaluate the benefits of predation by minute pirate bugs. Genetics Screening Cotton Genotypes for Reniform Nematode Resistance John E. Erpelding1, Salliana R. Stetina1 1Crop Genetics Research Unit, Agricultural Research Service, United States Department of Agriculture Here, a protocol is presented for the rapid non-destructive screening of cotton genotypes for reniform nematode resistance. The protocol involves visually examining the roots of nematode-infected cotton seedlings to determine infection response. The vegetative shoot from each plant is then propagated to recover plants for seed production. Environment Use of Principal Components for Scaling Up Topographic Models to Map Soil Redistribution and Soil Organic Carbon Xia Li1,2, Greg W. McCarty2 1Department of Geographical Sciences, University of Maryland, 2Hydrology & Remote Sensing Laboratory, Agricultural Research Service, United States Department of Agriculture Landscape processes are critical components of soil formation and play important roles in determining soil properties and spatial structure in landscapes. We propose a new approach using stepwise principal component regression to predict soil redistribution and soil organic carbon across various spatial scales. Environment Detection of Viruses from Bioaerosols Using Anion Exchange Resin Joshua W. Schaeffer1, Jeffrey C. Chandler2, Margaret Davidson1,3, Sheryl L. Magzamen1, Alma Pérez-Méndez4, Stephen J. Reynolds1, Lawrence D. Goodridge5, John Volckens6, Alan B. Franklin2, Susan A. Shriner2, Bledar Bisha7 1High Plains Intermountain Center for Agricultural Health and Safety, Department of Environmental and Radiological Health Sciences, Colorado State University, 2National Wildlife Research Center, Wildlife Services, Animal and Plant Health Inspection Service, United States Department of Agriculture, 3Western Sydney University, 4Leprino Foods, Inc, 5Department of Food Science and Agricultural Chemistry, McGill University, 6Department of Mechanical Engineering, Colorado State University, 7Department of Animal Science, University of Wyoming An anion exchange resin-based method, adapted to liquid impingement-based bioaerosol sampling of viruses is demonstrated. When coupled with downstream molecular detection, the method allows for facile and sensitive detection of viruses from bioaerosols. Environment Experimental Protocol for Biodiesel Production with Isolation of Alkenones as Coproducts from Commercial Isochrysis Algal Biomass Gregory W. O'Neil1, John R. Williams1, Julia Wilson-Peltier1, Gerhard Knothe2, Christopher M. Reddy3 1Department of Chemistry, Western Washington University, 2Agricultural Research Service, United States Department of Agriculture, 3Department of Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution Detailed methods are presented for the production of biodiesel along with the co-isolation of alkenones as valuable coproducts from commercial Isochrysis microalgae. Immunology and Infection Visualization of Twitching Motility and Characterization of the Role of the PilG in Xylella fastidiosa Xiangyang Shi1, Hong Lin2 1Department of Plant Science, University of California, Davis, 2Agricultural Research Service, San Joaquin Valley Agricultural Sciences Center, United States Department of Agriculture In this study, a nano-microfluidic flow chamber was employed to visualize and functionally characterize the twitching motility of Xylella fastidiosa, a bacterium that causes Pierce's disease in grapevine. Immunology and Infection Application of Long-term cultured Interferon-γ Enzyme-linked Immunospot Assay for Assessing Effector and Memory T Cell Responses in Cattle Mayara F. Maggioli1,2, Mitchell V. Palmer1, H. Martin Vordermeier3, Adam O. Whelan3, James M. Fosse4, Brian J. Nonnecke1, W. Ray Waters1 1Infectious Bacterial Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, 2Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, 3UK Veterinary Laboratories Agency, 4Visual Services, National Centers for Animal Health, Animal and Plant Health Inspection Service, United States Department of Agriculture Long-term cultured interferon-γ enzyme-linked immunospot assay is used as a measure of central memory responses and correlates with protective anti-mycobacterial vaccine responses. With this assay, peripheral blood mononuclear cells are stimulated with mycobacterial antigens and interleukin-2 for 14 days, enabling differentiation and expansion of central memory T cells. Environment Semi-High Throughput Screening for Potential Drought-tolerance in Lettuce (Lactuca sativa) Germplasm Collections Caleb Knepper1, Beiquan Mou1 1Agricultural Research Service, United States Department of Agriculture This protocol was developed to screen a large germplasm collection of the leafy vegetable lettuce (Lactuca sativa L.) for drought-tolerance, in order to identify a small candidate pool of lettuce for use in physiological, molecular, and genetic studies to identify underlying drought-tolerance traits along with breeding programs. Environment Preparation and Testing of Plant Seed Meal-based Wood Adhesives Zhongqi He1, Dorselyn C. Chapital1 1Southern Regional Research Center, Agricultural Research Service, United States Department of Agriculture To facilitate the effort in seeking more economic and environment-friendly formulations of natural product-based wood adhesives, this work demonstrates the preparation and testing of plant seed-based wood adhesives. This protocol allows one to assess plant seed-based agricultural products as suitable candidates for the substitution of synthetic-based wood adhesives. Biology Physiological Recordings and RNA Sequencing of the Gustatory Appendages of the Yellow-fever Mosquito Aedes aegypti Jackson T. Sparks1, Joseph C. Dickens1 1Agricultural Research Service, Henry A. Wallace Beltsville Agricultural Research Center, Plant Sciences Institute, Invasive Insect Biocontrol and Behavior Laboratory, United States Department of Agriculture Using two methods to estimate gene expression in the major gustatory appendages of Aedes aegypti, we have identified the set of genes putatively underlying the neuronal responses to bitter and repulsive compounds, as determined by electrophysiological examination. Chemistry Thin-layer Chromatographic (TLC) Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds Isabelle A. Kagan1, Michael D. Flythe1 1Forage-Animal Production Research Unit, Agricultural Research Service, United States Department of Agriculture Methods are described for thin-layer chromatographic (TLC) separation of plant extracts and contact bioautography to identify antibacterial metabolites. The methods are applied to the screening of red clover phenolic compounds inhibiting hyper ammonia-producing bacteria (HAB) native to the bovine rumen. Biology Utero-tubal Embryo Transfer and Vasectomy in the Mouse Model Pablo Bermejo-Alvarez1,2, Ki-Eun Park1,2, Bhanu P. Telugu1,2 1Animal Bioscience and Biotechnology Laboratory, United States Department of Agriculture, 2Department of Animal and Avian Sciences, University of Maryland Utero-tubal embryo transfer uses the utero-tubal junction as a barrier to prevent the embryo outflow that may occur when performing uterine transfer. Vasectomized males are required to obtain pseudopregnant recipients for embryo transfer. Both techniques are discussed.