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Submitted to: Journal of Visualized Experiments
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 6/24/2015 Publication Date: 1/28/2016 Citation: Hagler, J.R., Machtley, S.A. 2016. Administering and detecting protein marks on arthropods for dispersal research. Journal of Visualized Experiments. 107:e53693. doi: 10.3791/53693(2016). Interpretive Summary: Knowledge of insect movement patterns the nature is an important component of a successful pest management program. Arthropods are usually tracked by tagging them with a unique mark (colored dust or paints, tags, etc.) and then re-collecting them over time and space to determine their dispersal capabilities. Proteins have proven to be very effective for marking arthropods for ecological research. The proteins can then be detected on recaptured arthropods with highly sensitive and protein-specific assays. ARS scientists at Maricopa, AZ have pioneered the protein marking procedure. Here are described protocols for externally and internally tagging arthropods with protein. Then, a simple step-by-step guide is provided for the layman on how to detect the protein by two different types of immunological assays. In this demonstration, various aspects of the acquisition and detection of protein markers on arthropods for insect dispersal/migration research are discussed along with the various ways that the immunomarking procedure can be modified to suit a wide variety of research needs. Technical Abstract: Monitoring arthropod movement is often required to better understand their population dynamics, dispersal patterns, host plant preferences, and other ecological interactions. Arthropods are usually tracked in nature by tagging them with a unique mark and then re-collecting them over time and space to determine their dispersal capabilities. In addition to actual physical tags like colored dust or paint, various types of proteins have proven very effective for marking arthropods for ecological research. Proteins can be administered internally and/or externally. The proteins can then be detected on recaptured arthropods with a protein-specific enzyme-linked immunosorbent assay (ELISA). Here we describe protocols for externally and internally tagging arthropods with protein. Two simple experimental examples are demonstrated: (1) an internal protein mark introduced to an insect by feeding it a protein-enriched diet and (2) an external protein mark topically applied to an insect using a medical nebulizer. Then we relate a step-by-step guide of the sandwich and indirect ELISA methods used to detect the protein marks on the insects. In this demonstration, various aspects of the acquisition and detection of protein markers on arthropods for mark-release-recapture, mark-capture and self-mark-capture type research are discussed along with the various ways that the immunomarking procedure has been adapted to suit a wide variety of research protocols. |
